Gene profiling in CBA and BL/6 bone marrow derived dendritic cells (BMDC)
ABSTRACT: Analysis of baseline gene expression in bone marrow derived dendritic cells (BMDC) from female CBA/J (CBA) and C57BL/6 (BL/6) mice. Results provide insight into strain-dependent differences in gene expression. CBA and BL/6 BMDCs prepared from individual mice were plated in replicate at 1x10^6 cells/ml in 48-well tissue culture plates (BD Falcon). Replicates were pooled after 4hr and total RNA was obtained by Trizol® (Invitrogen) extraction according to the manufacturer’s instructions.
Project description:The Suppressor of cytokine signaling (SOCS) family of negative regulatory proteins are upregulated in response to several cytokines and pathogen-associated molecular patterns (PAMPs), and suppress cellular signaling responses by binding receptor phosphotyrosine residues. Exposure of bone marrow-derived dendritic cells (BMDCs) to 1D8 cells, a murine model of ovarian carcinoma, suppresses their ability to express CD40 and stimulate antigen specific responses in response to PAMPs, and in particular to poly I: C with the upregulated SOCS3 transcript and protein levels. The ectopic expression of SOCS3 in both the macrophage cell line RAW264.7 and BMDCs decreased signaling in response to both poly I:C and IFNα. Further, knockdown of SOCS3 transcripts significantly enhanced the responses of RAW264.7 and BMDCs to both poly I: C and IFNα. Immunoprecipitation and pull-down studies demonstrate that SOCS3 binds to the IFNα receptor TYK2. Since poly I: C triggers autocrine IFNα signaling, binding of SOCS3 to TYK2 may thereby suppress the activation of BMDCs by polyI:C and IFNα. Thus, elevated levels of SOCS3 in tumor-associated DCs may potentially resist the signals induced by TLR3 ligands and type I interferon to decrease DC activation via binding with IFNα receptor TyK2. Experiment Overall Design: Microarray analysis was used to compare the expression levels of Control mouse bone marrow-derived dendritic cells (BMDC), with cells that had been cocultured (1:5, tumor:BMDC) with mouse ovarian surface epithelial cell line (1D8) cells, with irradiated (50Gy) 1D8 cells, or with the supernatent from 1D8 cells (25%, v/v). Experiment Overall Design: One biologic sample was analyzed for each condition, four samples in all.
Project description:small RNAseq was preformed on Wt bone marrow-derived dendritic cells (BMDC) and miR-155 and miR-146a double knockout (DKO) BMDCs that received Wt exosomes to investigate the differences in transferred miRNA Small RNA profiles were generated from Wt donor BMDCs and DKO BMDCs given Wt exosomes 3 replicates in each group
Project description:Innate immunity is the first line of defense against viral and microbial pathogens. BMDC is critical for innate immunity. To investigate the complicated net signaling after virus invasion, we did a cDNA microarray analysis of BMDC with or without Sendai Virus infection. We used microarrays to find proteins that upregulated by Sendai Virus infection and investigated if these proteins had functions in regulating Sendai Virus induced signaling pathway. BMDC cells are seperated from C57BL6 mice, infected with Sendai Virus or not,cultured and harveseted for RNA extraction and hybridization on Affymetrix microarrays
Project description:Previously we have shown significant differences in lactation performance, mammary gland histology and expression profiles of mammary transcriptome during peak-lactation (lactation day 9; L9) between the ordinary CBA/CaH (CBA) and the superior QSi5 strains of mice. In the present study, we compared mammary gland histology between CBA and QSi5 at mid-pregnancy (pregnancy day 12; P12). We assessed lactation performance during the first 8 days of lactation of the 13th - 14th generation of the Advanced Intercross Line (AIL) (CBA X QSi5) mice. We utilized an integrative approach to analyzing mammary microarray expression profiles of CBA and QSi5 at P12 and CBA, AIL and QSi5 at L9. The inguinal mammary glands of CBA/CaH and QSi5 during mid-pregnancy (Pregnancy day 12; P12), and the glands of CBA/CaH, AIL and QSi5 during peak lactation (Lactation day 9; L9) were collected and total RNA was extracted for Affymetrix microarray (mouse genome 430 2) assay
Project description:The gene expression difference between CBA/J and DBA/2 mice after the injection of CAWS (Candida albicans water-soluble fraction) was compared using an oligo-DNA microarray. Overall design: CAWS was injected into CBA/J and DBA/2 mice. 21 days after the injection, RNA was extracted from the splenocytes and measured using the mouse oligo-DNA 10K micoarray.
Project description:mouse primary BMDCs were stimulated with tlr ligands and gene expression changes were profiled on Affymetrix arrays BMDC were stimulated with 5 tlr ligands (LPS, pIC ,PAM, CpG, GRD) across 9 time points (.5, 1, 2, 4, 6, 8, 12, 16, 24 hours). Unstimulated cells were used as controls.