intima. Here, they receive a series selleck bio of differentiation signals including macrophage col ony stimulating factor and minimally o idized LDL that enables them to mature into macrophages. These macrophages then engulf large quantities of cholesterol to become lipid laden foam cells. And it is the accumulation of these foam cells that eventually leads to the formation of characteristic fatty streaks, intermediate lesions and fibrous plaques. To date, though, the actual role of chemokines and their receptors in atherosclerosis has not been clearly estab lished. However, recent studies using transgenic mouse models of atherosclerosis have provided convincing evi dence that CCR2 is required for disease progression in apolipoprotein E null mice.
In these animals, dis ruption of the CCR2 gene greatly decreases lesion forma tion without affecting plasma lipid or lipoprotein concentrations. Using a slightly different approach Roll ins and colleagues have demonstrated that CCL2, the lig and for CCR2, plays an equally important role in the development of atherosclerosis in low density lipoprotein receptor deficient mice. Here, deletion of CCL2 leads to a significant reduction in lipid deposition within the aorta. Despite the promising e perimental results from these systems, relatively little is known about how the e pres sion of chemokine receptor genes is regulated in normal or diseased human tissues. A recent paper by Yamamoto and colleagues e amined the basal regulatory mech anisms underlying e pression of the CCR2 gene in the human monocyte cell line, THP 1.
Indeed, this group characterized two key elements that seemed to be neces sary and sufficient for the basal regulation of CCR2 e pression an Oct 1 binding sequence located 36 bp upstream of the TATA bo and a tandem CAAT enhancer binding protein binding sequence located, unu sually, in the 5 UTR. However, studies have not directly e amined the molecular mechanisms by which basal e pression of CCR2 is rapidly downregulated during the differentiation of monocytes into macro phages. In an effort to address this issue, we have further devel oped a model of monocyte differentiation using THP 1 cells, which can be induced to mature into macrophages using either phorbol esters and ionomycin or a physiolog ical combination of interferon and M CSF.
AV-951 In common with other studies, we report here that THP 1 cell maturation mediated by either high concentrations of PMA alone, or very low concentrations of PMA plus ionomycin is characterized by an increase in size, the development of an adherent pheno type new product and the up regulation of a panel of differentiation markers, in addition, CCR2, but not CCR1, was specifically down regulated during differentiation. Modu lation of CCR2 by PMA, but not PMA plus ionomycin, was found to be sensitive to inhi bition by the broad spectrum protein kinase inhibitor staurosporine. Furthermore, transient transfection of THP 1 cells with a CCR2 specific reporter construct indi cated that PM
poly morphism of pre miR 196a2 gene was observed in several malignant diseases, including head and neck cancer, and has been associated with cancer susceptibility or prognosis. These studies indicate that miR 196 dysregulation plays an important role in carcinogenesis. Consistent selleck chem with other reports, we previously observed miR 196 overe pression in oral cancer cell lines. In that study, we fur ther identified miR 196 overe pression in the cancer tissues of appro imately 90% of patients with oral cancer compared to their e pression in normal tissue, and this overe pression was associated with an aggressive phenotype with lymph node invasion. These re sults demonstrate the significance of miR 196 in the devel opment of oral cancer. There are limited reports on the role of miR 196 in cancer development.
In a study of breast cancer, ectopic miR 196a e pression suppressed cell invasion, but the e pression level of miR 196 was not correlated with the clinical metastatic status. This result is different from the findings in gastrointestinal cancers, in which miR 196 overe pression promotes cell migration and in vasion in colorectal and gastric cancer cells. In another study, transfection of miR 196a mimic oligo nucleotides into esophagus cells revealed that miR 196a promoted cell proliferation and suppressed apoptosis. Thus far, the function of miR 196 in cancer re mains obscure. In this study, we determined the role of miR 196 in oral cancer using both silencing and overe pression approaches.
We found that both miR 196a and miR 196b actively promoted cell migration and invasion, which were supported by the altered e pression of fibronectin and N cadherin. However, neither miR 196a nor miR 196b affected cell growth and colony formation. Hence, the main function of the miR 196 family in oral cancer is the regulation of cell mobility and invasiveness. Identification of the target molecules of these miRNAs is of high interest. Previously, several molecules were re ported as regulatory targets of miR 196. HO family members have been reported primarily, including HO B7 in melanoma, HO A in acute lymphoblastic leukemia, and HO C8 in melanoma and breast cancer. In this study, four HO family genes were identified as targets of miR 196. However, none of these genes responded to miR 196 perturbation. This difference may be due to the distinct tissue specificity among the various regulatory targets of miR 196.
In this study, after clarifying the correlation between NME4 and miR 196 e pression in cells, assessing the response of NME4 to miR 196 modulation, we identified NME4 as a direct target of miR 196a and miR 196b in oral cancer Entinostat using a luciferase reporter assay, NME4, also named nm23 H4, is a member of the nm23 family. The proteins in this family possess nu cleoside diphosphate kinase activity, which is believed to be involved in DNA repair mechanisms. Nm23 fam ily proteins also contain the RGD domain, as they can bind to integrin, namely and this family has been post
vity of T47D cells to do orubicin, etoposide and camptothecin, sug gesting that the activation of NF B cIAP2 signaling pathway by retinoids in these cells correlates with an increase in apoptosis resistance. On the other hand, in H3396 cells where 9 cis RA induces neither NF B http://www.selleckchem.com/products/Axitinib.html acti vation nor cIAP2 e pression but makes the cells enter a fully apoptotic program, death curves showed that the treatment with 9 cis RA not only induced apoptosis by itself, but also increased in an additive manner the apoptosis in response to TRAIL, etoposide, do orubicin or camptothecin. Note that, 9 cis RA treatment augmented apoptosis mediated by the death receptor pathway in both cell lines. These results reveal that the activation of NF B cIAP2 signaling pathway by reti noids in a given breast cancer cell apparently correlates with the ability of these retinoids to protect cells against chemotherapy induced apoptosis.
We further investigated the effect of 9 cis RA in pre venting etoposide mediated apoptosis in one additional breast cancer cell line, ZR 75 1, where the retinoid upregulates cIAP2 e pression and potentially NF B activation. ZR 75 1 cells responded to 9 cis RA in a similar manner to T47D cells showing a reduction in sensitivity to etoposide upon pretreat ment with 9 cis RA. These results show that the protection against etoposide mediated cell death e erted by 9 cis RA is not restricted to T47D breast cancer cells. cIAP2 is not critically involved in the protection of etoposide induced apoptosis by 9 cis RA Previous reports have shown that e ogenous overe pres sion of cIAP2 can abrogate apoptosis induced by geno to ic anticancer drugs such as etoposide, but not death receptor mediated apoptosis in a NF B null back ground.
We have observed that 9 cis RA protection against etoposide mediated cell death correlates with cIAP2 induced e pression by 9 cis RA in T47D cells. To test whether cIAP2 has a role in this protection, e pression of cIAP2 was suppressed by using transiently e pressed siRNA. Although the cIAP2 siRNA efficiently downregulated both basal and 9 cis RA induced Dacomitinib cIAP2 protein levels when compared to a scrambled siRNA, downregulation of cIAP2 was not sufficient to restore sensitivity to etoposide mediated cell death in 9 cis RA pretreated T47D cells.
To further con firm the above results, we compared the level of activa tion of caspase 3 by western blot, as a measurement of cell death, between find more info T47D cells transfected with a scrambled siRNA and T47D cells transfected with a siRNA against cIAP2. While the levels of cleaved cas pase 3 were induced by etoposide and strikingly abro gated when cells were pretreated with 9 cis RA in scrambled siRNA transfected cells, downregulation of cIAP2 protein level did not restore the levels of cleaved caspase 3 induced by etoposide in the presence of 9 cis RA. This reveals that cIAP2 is not critically involved in the inhibition of etoposide induced apopto sis in 9 cis RA pretreated T47D cells. Over e pression o
RNeasy Mini kit following the manufac turers protocol for plant tissues. cDNA AFLP analysis We used the cDNA AFLP protocol described by Vos et al. and Bachem et al. with the modifications and primers described by Breyne et al. which per selleck Olaparib mit the visualization of a single cDNA fragment for each mRNA present in the original sample, reducing the output sequence redundancy. Double stranded cDNA was synthesized from 2. 5 ug total RNA using the Super script II reverse transcription kit and a bio tinylated oligo dT primer. After pre amplification, the mixture was diluted 600 fold and 5 ul was used for selective amplification with 128 primer combinations, carried out with one selective nucleotide added on the 33P labeled BstYI primer and two selective nucleotides on the MseI primer.
We used the following touch down PCR conditions, 2 min dena turation at 94 C followed by 30 s denaturation at 94 C, 30 s annealing at 65 C, 60 s extension at 72 C for 13 cycles, 30 s denaturation at 94 C, 30 s annealing at 56 C, 60 s extension at 72 C for 23 cycles, and 5 min at 72 C. Selective ATP labeled amplification products were separated on a 6% polyacrylamide gel in a Sequi Gen GT Sequencing Cell running for 2. 5 h at 115 W and 50 C. Gels were dried onto 3 MM Whatman paper for 2 h at 80 C on a Gel Dryer Model 583 and marked with Glogos II Autorad Markers before exposing to Kodak Biomax MR films, for 12 16 h.
Sequence analysis of cDNA AFLP fragments Bands corresponding to differentially expressed genes were excised from the gels with a surgical blade and the eluted DNA was reamplified using non labeled primers identical to those employed for selective amplification and the following PCR conditions, 15 min denaturation at 94 C, 40 s denaturation at 94 C, 60 s annealing at 56 C, 40 s extension at 72 C for 35 cycles, and 5 min at 72 C. The quantity of each reamplified bands were checked on a 1. 8% agarose gel against the 1650 bp fragment of the DNA ladder 1 Kb plus. PCR products were purified with MultiScreen PCR u96 plates and sequenced directly. Sequence information was obtained by comparing nucleotide and protein sequences in the available public databases by BLAST sequence alignment. Homol ogy searching was carried out against the following data bases, NCBI, Cucurbit Genomic Database Melon Unigene ver. 4. 0, AV-951 UNIPROT database and Fusarium Comparative Database.
Sequences were manually assigned to functional categories based on the analysis of scientific literature and also with the aid of the information reported for each sequences by the Gene Ontology Consortium, where available. Sequence data from moreover this article have been deposited in GenBank, Accession Numbers, HO867279 HO867981. Real time RT PCR analysis Real time RT PCR was carried out on pools of RNA derived from two independent biological experiments. All samples were examined as three technical repli cates. Samples were prepared from whole stems of infected and mock inoculated plants correspond ing, for each gene te
1. 5 mTorr. Samples were analyzed using selected reaction monitoring mode with a scan width selleckchem of 1 m z and a scan time of 0. 05 s. The SRM parameters for most metabolites have been published previously. This method was used to scan for almost 300 meta bolites. Xcalibur software was used to manually assess the elution time of the correct LC spectral peak for each metabolite specific SRM. The Quan Browser utility in Xcalibur was then used to integrate the LC spectral peak area for each detected compound, and these data were exported to a Microsoft Excel spreadsheet for fur ther processing. Statistical analysis Statistical analysis of the microarray data was performed using R 2. 9. 0 and routines contained in Bioconductor. GC robust multi array average was used to normalize and scale the raw data from CEL files.
The normalized data were filtered for low expression by removing any probes with normalized expression less than 3 in at least 5 arrays. Statistical significance of gene expression differences were analyzed by one way ANOVA and empirical bayes using the limma package. Differential expression was defined based on false discovery rate adjusted p value 0. 05. False discovery rate for differential expression and for GO and KEGG enrichment testing was controlled using the Benjamini Hochberg method. Venn diagrams of differentially expressed genes were plot ted to visualize the number of differentially expressed genes for each treatment comparison and their intersec tions. Hierarchical clustering of significant genes was per formed using the hclust function and a hierarchical clustering heatmap was created using heatmap.
2 in the gplots package. Hierarchical clustering also was used to identify correlated patterns of gene expression and meta bolites. The Database for Annotation, Visualization and Integrated Discovery and ClueGO, a Cytoscape plug in, were used for Gene Ontology at level 6 and 7 and KEGG analysis of differentially expressed genes. Statistical analysis of metabolomic data was performed using an analysis tool that we developed specif ically for metabolomic data analyses. The script, written in the language R, uses linear mixed effect modeling to normalize metabolomics data AV-951 containing both fixed and random effect confounding variables. The script averages any replicate measurements made on ex perimental units and performs ANOVA to test for statis tical differences between experimental groups.
Aquaculture is the fastest growing animal production activity worldwide, supplying an increasing proportion of BAY 734506 fish for human consumption, estimated at around 50% of total supply in 2008. However, the growth of marine aquaculture is threatened by its excessive reli ance on fishmeal and fish oil from wild stocks for the production of fish feeds, which is also an eco logically unsound practice. Almost 89% of the total glo bal production of FO is currently used by aquaculture and the future of this activity strongly depends on the reduction of dependen
and showing selleck chemical the largest fold change, we also performed qRT PCR using cerebellum tissue samples from 10 PGRN and 30 PGRN FTLD TDP patients, using simi lar techniques and analyses, as described above. Indivi dual Ct values in the validation data sets for both the cortex and cerebellum are provided in Additional File 5. miRNA expression in vitro SH SY5Y cells were cultured in a 1,1 mixture of OPTI MEM and DMEM containing 5% heat inactivated FBS and 1% penicillin streptomycin. Cell cultures were kept at 37 C in a humidified atmosphere with 5% CO2. Cells were seeded at 175, 000 cells well in 6 well plates and 24 h later transfected with siRNA against PGRN or negative control siRNA at a final concentration of 25 nM using Lipofectamine2000.
After 48 h of transfection, total RNA was extracted from SH SY5Y cells and quantitative RT PCR was performed as described above in the miRNA validation methods section. Bioinformatics analysis It has been extensively reported that miRNAs primarily decrease mRNA expression and repress translation. For the miRNA candidates significantly dysregulated in the frontal cortex and cerebellum of PGRN FTLD TDP patients, we identified their predicted mRNA targets through TargetScan. For each of those miRNAs, we com pared their predicted gene targets with mRNA expression results of PGRN and PGRN FTLD TDP patients depos ited in the Gene Expression database. The GEO Affy metrix array dataset published by Chen Plotkin et al. profiled mRNA levels in several tissue types from controls, as well as PGRN and PGRN FTLD patients.
The significantly dysregulated miRNAs which were anti correlated in expression with their mRNA targets in the Affymetrix data set were further ana lyzed by Ingenuity software for insight into their biological roles. Aspergillus niger is a ubiquitous ?lamentous fungus. According AV-951 to its saprophytic lifestyle, A. niger is capable of secreting large amounts of various plant polysaccharide degrading enzymes. Its naturally high secretion capacity has long been exploited in industrial biotechnology for the production of homologous and heterologous proteins as well as organic acids. Many of its products have acquired the GRAS status, meaning that they are gen erally considered as safe food ingredients. However, besides its positive economic relevance as an industrial workhorse, A.
niger is a common storage mold causing spoilage of agricultural goods and contamination of food and feedstocks with mycotoxins. Although to a much lesser extent than other species of its genus, A. niger is an opportunistic pathogen, which can cause invasive aspergillosis in immunocompromised patients. A. niger is exclusively known to propagate via an asex ual life cycle, selleck chemicals llc which ?nally leads to the formation of black airborne mitotic spores. Core genes involved in signal transduction and conidiophore development in the model fungus A. nidulans have also been identi?ed in A. niger, suggesting that the regulation of asexual development is conserved
This agrees with the result from metabolism studies with compound 1 that the primary route of biotransformation is oxidation, mainly at the para position of the phenyl ring and the a 17-AAG CAS position of the sulfonyl group in the aliphatic side chain.
Isothermal titration calorimetry (ITC) was used to investigate the binding of six inhibitors to 1-deoxy-Dxylulose-5-phosphate reductoisomerase (DXR), a target for developing novel anti-infectives. The binding of hydroxamate inhibitors to Escherichia coli DXR is Mg2+-dependent, highly endothermic (Delta H, 22.7-24.3 kJ/mol), and entropy-driven, while that of nonhydroxamate compounds is metal ion-independent and exothermic (Delta H, -19.4 to -13.8 kJ/mol), showing that hydration/dehydration of the enzyme metal ion binding pocket account for the drastic Delta H change.
However, for DXRs from Plasmodium falciparum and Mycobacterium tuberculosis, the binding of all inhibitors is exothermic (Delta H, -24.9 to -9.2 kJ/mol), suggesting that the metal ion binding sites of these two enzymes are considerably less hydrated. The dissociation constants measured by ITC are well correlated with those obtained by enzyme inhibition assays (R-2 = 0.75). Given the rapid rise of antibiotic resistance, this work is of interest since it provides novel structural implications for rational development of potent DXR inhibitors.
Toll-like receptors (TLRs) are key targets in the design of immunomodulating agents for use as vaccine adjuvants and anticancer treatments. The imidazoquinolines, imiquimod and resiquimod, have been shown to activate TLR-7 and -8, which in turn induce cytokine production as part of the innate immune response.
Herein, we report the synthesis and discovery of a C7-methoxycarbonyl derivative of imiquimod that stimulates cytokine production but is devoid of TLR-7/8 activity. Data AV-951 are presented that shows that this analogue not only induces IL-12p40 and TNF production, similar to that of imiquimod and resiquimod, but greatly enhances the production of IL-1 beta, a key cytokine involved in the activation of CD4 T cells. It is further demonstrated that TLR-7/8 activation can be recovered by the addition of a C2-alkyl substituent to this newly discovered analogue. The results support the existence of an alternative mechanism of action by which imidazoquinolines can stimulate cytokine production.
Natural and synthetic histone deacetylase (HDAC) inhibitors generally derive their strong binding affinity and high potency from a key functional group that binds to the Zn2+ ion within the enzyme active site. However, this feature is also thought to carry the potential liability of undesirable off-target interactions with other metalloenzymes. As a step toward mitigating this issue, selleck here, we describe the design, synthesis, and structure-activity characterizations of cyclic alpha(3)beta-tetrapeptide HDAC inhibitors that lack the presumed indispensable Zn2+-binding group. The lead compounds (e.g.
Its unique structure also imparts chemical function that allows it also to mediate charge transport (CT). We have utilized diverse platforms selleck kinase inhibitor to probe DNA CT, using spectroscopic electrochemical, and even genetic methods. These studies have established powerful features of DNA CT chemistry. DNA CT can occur over long molecular distances as long as the bases are well stacked. The perturbations in base stacking that arise with single base mismatches, DNA lesions, and the binding of some proteins that kink the DNA all inhibit DNA CT. Significantly, single molecule studies of DNA CT show that ground state CT can occur over 34 nm if the duplex is well stacked; one single base mismatch inhibits CT. The DNA duplex is an effective sensor for the Integrity of the base pair stack.
Moreover, the efficiency of DNA CT is what one would expect for a stack of graphite sheets: equivalent to the stack of DNA base pairs and independent of the sugar-phosphate backbone.
Since DNA CT offers a means to carry out redox chemistry from a distance, we have considered how this chemistry might be used for long range biological signaling We have taken advantage of our chemical probes and platforms to characterize DNA CT in the context of the cell. CT can occur over long distances, perhaps funneling damage to particular sites and insulating others from oxidative stress. Significantly, transcription factors that activate the genome to respond to oxidative stress can also be activated from a distance through DNA CT.
Numerous proteins maintain the integrity of the genome and an increasing number of them contain [4Fe-4S] clusters that do not appear to carry out either structural or enzymatic roles. Using electrochemical methods, we find that DNA binding shifts the redox potentials of the dusters, activating Dacomitinib them towards oxidation at physiological potentials.
We have proposed a model that describes how repair proteins may utilize DNA CT to efficiently search the genome for lesions. Importantly, many of these proteins occur in low copy numbers within the cell, and thus a processive mechanism does not provide a sufficient explanation of how they find and repair lesions before the cell divides. Using atomic force microscopy and genetic assays, we show that repair proteins proficient at DNA CT can relocalize in the vicinity of DNA lesions and can cooperate in finding lesions within the cell. Conversely, proteins defective in DNA CT cannot relocalize Rapamycin AY-22989 in the vicinity of lesions and do not assist other proteins involved in repair within the cell. Moreover such genetic defects are associated with disease in human protein analogues.
The difference in MtLs between pediatric and adult MDS might be related to the physiological hypermethylation of tumor suppressor genes in aging. Copyright (C) 2013 S. Karger AG, Basel
Homocysteine is an amino acid, which plays several important roles in human physiology. A wide range of disorders, including neuropsychiatric selleckchem disorders and autism, are associated with increased homocysteine levels in biological fluids. Various B vitamins: B6 (pyridoxine), B12 (cobalamin), and B9 (folic acid) are required as co-factors by the enzymes involved in homocysteine metabolism. Therefore, monitoring of homocysteine levels in body fluids of autistic children can provide information on genetic and physiological diseases, improper lifestyle (including dietary habits), as well as a variety of pathological conditions.
This review presents information on homocysteine metabolism, determination of homocysteine in biological fluids, and shows abnormalities in the levels of homocysteine in the body fluids of autistic children.
Inflammation is a non-specific immune response to infection, irritation or other injury, the key features being redness, warmth, swelling and pain. A number of mediators are released which alter the resistance of mucosa to injury induced by noxious substances. Oxidative stress is a unifying mechanism of injury in many types of disease processes, including gastrointestinal diseases. It has been defined as an imbalance in the activity of pro and antioxidants. Pro-oxidants favour free radical formation while antioxidants inhibit or retard the same.
A number of markers of oxidative stress have been identified. This review provides an overview of various mediators of inflammation and oxidative stress, and diverse approaches for prevention and treatment of gastrointestinal inflammation.
Glycosylation is the most common chemical process of protein modification and occurs Brefeldin_A in every living cell. Disturbances of this process may be either congenital or acquired. Congenital disorders of glycosylation (CDG) are a rapidly growing disease family, with about 50 disorders reported since its first clinical description in 1980. Most of the human diseases have been discovered recently. CDG result from defects in the synthesis of the N- and O-glycans moiety of glycoproteins, and in the attachment to the polypeptide chain of proteins.
www.selleckchem.com/products/Sunitinib-Malate-(Sutent).html These defects have been found in the activation, presentation, and transport of sugar precursors, in the enzymes responsible for glycosylation, and in proteins that control the traffic of component. There are two main types of protein glycosylation: N-glycosylation and O-glycosylation. Most diseases are due to defects in the N-glycosylation pathway. For the sake of convenience, CDG were divided into 2 types, type I and II. CDG can affect nearly all organs and systems. The considerable variability of clinical features makes it difficult to recognize patients with CDG.
To determine whether over expression of TBX3 affects mammary stem cell proliferation, we performed FACS analysis of the stem like cell population, Lin CD24 mice and their un induced littermate controls. We example found that over expression of TBX3 significantly increased the frequency of Lin CD24 CD29high stem like cell population, indicating that TBX3 expression is associated with an increased number of mammary stem like cells. This could explain another mechanism by which TBX3 over expression can cause hyperplasia and accelerated mammary gland develop ment. Further studies of the mechanisms by which TBX3 regulates mammary stem like cells are required to improve our understanding of mammary gland devel opment and TBX3 function.
Conclusions TBX3 over expression causes mammary gland hyperpla sia possibly by inhibiting NF BIB expression and thus promoting cell proliferation. Also, over expression of TBX3 is associated with an increased number Dacomitinib of mam mary stem like cells suggesting another mechanism by which TBX3 may promote mammary gland hyperplasia and contribute to breast cancer development. Methods Plasmid construction To generate the Tet on inducible N myc TBX3 expres sion cassette, the full length human TBX3 cDNA fused with the N myc tag was subcloned from the expression vector, pcDNA myc TBX3, into the ClaI and SpeI sites of the TMILA plasmid, downstream of an inducible tetracycline pro moter. Correct insertion of the N myc TBX3 transgene into the TMILA plasmid was verified by sequencing.
Generation and PCR genotyping of transgenic mice To generate doxycycline inducible myc TBX3 transgenic mice, the N myc TBX3 expression cassette was cut out from the TMILA myc TBX3 plasmid using the PvuII restriction enzyme to remove the plasmid backbone. The fragment was gel purified using the Qiagen Gel Extraction Kit and filtered using a 0. 1 micron filter. The purified DNA fragment was then diluted with injection buffer to a 2ng ul concentration and microinjected at the UCI Transgenic Mouse Facility. A total of 176 fertilized eggs were injected. expression cassette were used as founders to cross with established MMTV rtTA mice to create double trans genic mice. Doxycycline administration Transgene expression was induced by adding 2 mg ml doxycycline to the drinking water from weaning age as previously described. All mice involved in the experiments were examined weekly for palpable tumor formation.
In vivo imaging of Tet on inducible TBX3 luciferase reporter system For in vivo mouse imaging, a cooled ICCD camera was placed on top of a light tight box. Prior to imaging, mice were Erlotinib FDA sedated by intraperitoneal injection of 250 ng Xylazine and 2 mg Ketamine. After 5 minutes, an aqueous solution of luciferin was injected into the peritoneal cavity at 150 mg kg body weight.