Project description:Unstimulated murine bone marrow derived macrophages cultured in L92 media from WT (4 biological replicates), Nrf2 KO (3 biological replicates) and Keap1 KD BMDM (3 biological replicates) were processed and analysed utilising DIA (label free) proteomic analysis. The Nrf2 KO mouse (DOI: 10.1006/bbrc.1997.6943 ) and Keap1 KD mouse ( DOI: 10.1128/MCB.01591-09) were previously published as noted.
Project description:murine p16ink4a deficient (p16ko) and control (p16wt) bone marrow cells were either differentiated with normal LCM-supplemented differentiation medium to obtain bone marrow derived macrophages (BMDM) or supplemented with Interleukin 4 during differeniation to obtain M2 polarized p16wt and p16ko BMDM.
Project description:Purpose: Next-generation sequencing (NGS) has revolutionized systems-based analysis of cellular pathways. The goals of this study are to evaluate the gene expression of M2-polarized bone marrow-derived macrophages (BMDM) compared with M0-BMDMs from wild type (WT) mouse. Methods: BMDMs were obtained from WT mice and polarized toward M2-BMDMs using IL-4 and M-CSF. M0-BMDMs were maintained in culture with M-CSF only. Total RNA of M2- and M0-BMDMs was extracted. BMDM RNA profiles were generated by deep sequencing for two groups (M2- versus M0-BMDM) with three mouse samples each. Results: There were significant differences between M2- and M0-BMDMs. Conclusions: Polarization of BMDMs from M0 to M2 induces various changes at the transcription level.
Project description:Interferon regulatory factor 2 binding protein 2 (Irf2bp2) is a corepressor of Irf2 that suppresses inflammatory gene expression and promotes the M2 macrophage phenotype in bone marrow derived macrophages (BMDM). Loss of Irf2bp2 in male macrophages promotes an inflammatory phenotype. Here, we compared the expression profile of female BMDM to their male counterparts to identify genes differentially affected by the loss of Irf2bp2 by sex in primary macrophages cultured under basal conditions.
Project description:Bone marrow cells were isolated, primed with M-CSF (M-BMDM) or GM-CSF (GM-BMDM) and cultured for 7 days. The proteomic difference between GM-BMDM and M-BMDM were analyzed to describe the phenotye and function of two types of macrophages.
Project description:We reported the function of Roquin-1 in the miRNA-sorting of macrophages derived exosomes. At first, we used the supernatant of 929 cells to culture the bone marrow derived macrophages (BMDM) from bone marrow cells of WT and Roquin-1 san:san mice. Then, we isolated the macrophages derived exosomes by ultracentrifugation. At last, we performed Next-generation sequencing to detect the differences of miRNA-sorting between WT and Roquin-1 macrophages derived exosomes.
Project description:Introduction: HGFL-Ron signaling is augmented in human breast cancer and is associated with poor overall prognosis. Here, we investigate the role of HGFL-Ron signaling in RON-modulated murine macrophages through RNA-sequencing of bone marrow-derived macrophages from FVB WT or FVB RON tyrosine kinase -/- mice. BMDM of each genotype M2-polarized via 72 hour treatment with IL-4 were submitted for transcriptomic characterization on the Illumina HiSeq 2500. High quality reads were aligned to the mm10 genome and quantified to generate RPKM
Project description:Several members of the matrix metalloproteinase (MMP) family control specific immune processes, such as leukocyte influx and chemokine activity. Stromelysin-2 (MMP10) is expressed in numerous tissues after injury; however, little is known of its function. Here, we report that MMP10 is expressed by macrophages in human lungs from patients with cystic fibrosis and induced in mouse macrophages in response to Pseudomonas aeruginosa infection both in vivo and in cultured bone marrow-derived macrophages (BMDM). Whereas all wildtype mice were alive at 48 h post-infection, all Mmp10â/â mice had greater weight loss and died or reached euthanasia criteria with no defect in bacterial clearance. Rather, macrophage numbers in infected Mmp10â/â lungs were about 3-4-fold greater than in wildtypes. Demonstrating that the protective effect of MMP10 was due its production by macrophages, adoptive transfer of wildtype BMDM normalized infection-induced weight loss in Mmp10â/â recipients to wildtype levels. In vivo, the expression of several M1 macrophage markers were elevated and M2 markers reduced in Mmp10â/â lungs, and we saw similar differences between M1-activated wildtype and Mmp10â/â BMDM. Global gene expression analysis revealed that infection-mediated transcriptional changes persisted in Mmp10â/â BMDM long after they downregulated in wildtype cells. These results indicate that MMP10 serves a beneficial role in response to infection by moderating the pro-inflammatory response of infiltrating macrophages. Total RNA from bone marrow-derived macrophages from 3 WT and 3 Mmp10-/- mice for each treatment: media change only, 6h post PA infection or 24 h post PA infection. Samples were hybidized to 18 Affymetrix GeneChip Mouse Gene 2.0 ST arrays.
