Project description:We used microarrays to compare gene expression changes in macrophages caused by deletion of bcl6. Keywords: BCL6 deletion compared to wild-type Primary bone marrow-derived macrophages were isolated from C57BL/6 mice transplanted and reconstituted with wild type or bcl6 knockout bone marrow.
Project description:Using microarrays, we compared the changes in levels of gene expression between wild type mouse bone marrow derived macrophages upon treatment with the Bcl6 peptide inhibitor, RI-BPI, that specifically blocks interaction between BCL6 and the co-repressors NCoR or SMRT. Total RNA was obtained from cultered wild type primary bone marrow-derived macrophages that were treated with either 5 μM control or RI-BPI peptide in MSF media for 12 hours.
Project description:Using ChIP-seq, we reveal the SMRT and NCoR co-repressor cistromes, which each consist of over 30,000 half-shared binding sites. Moreover, we identify Bcl6-bound sub-cistromes for each co-repressor, which are strongly concentrated on NF-κB-driven inflammatory and tissue remodeling genes. These results reveal a critical role for Bcl6 and its corepressors SMRT and NCoR in the prevention of atherosclerosis and chronic inflammation. Identification of SMRT and NCoR binding sites in wild-type and Bcl6 knockout primary bone-marrow derived macrophages
Project description:Using microarrays, we compared the changes in levels of gene expression between wild type mouse bone marrow derived macrophages upon treatment with the Bcl6 peptide inhibitor, RI-BPI, that specifically blocks interaction between BCL6 and the co-repressors NCoR or SMRT.
Project description:Using microarrays, we compared the changes in levels of gene expression between wildtype and Bcl6 KO macrophages in the absence or presence of LPS. Total RNA was obtained from WT and Bcl6 KO unstimulated and LPS-stimulated primary bone marrow-derived macrophages
Project description:Bone marrow-derived macrophages were generated from wild type and Junb knockout mice and tested for their responses to LPS treatment. Independent replicate wells from wild type and knockout bone marrow-derived macrophages were mock-treated or LPS-treated. RNA was isolated at 4 h post-treatment, amplified, and hybridized to Agilent Mouse 8x60k microarrays as a single color experiment.
Project description:To identify factors that could explain why mice transplanted with Vim deficient bone marrow display decreased atherosclerosis despite increased inflammation, we performed global gene expression profiling of bone-marrow derived macrophages from vimentin-deficient or wild-type littermates on C57BL/6 background.
Project description:Bone marrow-derived macrophages were generated from wild type and Junb knockout mice and tested for their responses to LPS treatment.
Project description:RNA-sequencing of uninfected and Moraxella catarrhalis-infected bone marrow-derived macrophages (BMDMs) isolated from Ifnar1 knockout and wild-type mice.
Project description:Previous work has shown conflicting roles for Tec family kinases in regulation of Toll-like receptor (TLR)-dependent signalling in myeloid cells. In the present study, we performed a detailed investigation of the role of Btk and Tec kinases in regulating TLR signalling in several types of primary murine macrophages. We demonstrate that primary resident peritoneal macrophages deficient for Btk and Tec secrete less pro-inflammatory cytokines in response to TLR stimulation than wild type cells. In contrast, we found that bone marrow-derived and thioglycollate-elicited peritoneal macrophages deficient for Btk and Tec secrete more pro-inflammatory cytokines than wild type cells. We then compared the phosphoproteome regulated by Tec kinases and lipopolysaccharide in primary peritoneal and bone marrow derived macrophages. From this analysis we determined that Tec kinases regulate different signalling programs in these cell types. In additional studies using bone marrow-derived macrophages, we find that Tec and Btk promote phosphorylation events necessary for immunoreceptor-mediated inhibition of TLR signalling. Taken together, our results are consistent with a model where Tec kinases (Btk, Tec, Bmx) are required for TLR-dependent signalling in many types of myeloid cells. However, our data also support a cell type-specific TLR-inhibitory role for Btk and Tec that is mediated by immunoreceptor activation and signalling via PI3K.
