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:Macrophages show substantial plasticity, leading to a diverse population of these cells with different states of polarization during differentiation from bone marrow. However, the mechanistic insights into this process are not well understood. Here, we identified a novel role of ribosomal protein L13a previously shown to be engaged in the physiological control of inflammation regulating macrophage diversity and polarity. Using an ex-vivo differentiation model of bone marrow-derived macrophages (BMDM) from the control (L13aflox/flox) and myeloid-specific L13a KO (L13aflox/flox LysMCre+) mice (L13a-KO) we presented compelling evidence of the role of L13a in regulating macrophage polarization that goes beyond the M1-M2-based binary concept. We show that macrophages from L13a-KO mice lead to enhanced expression of classical markers of both M1 and M2 and surprising deviation from the expected response under known inducers of polarity. The phosphorylation-dependent activation of a number of signaling molecules played a role in this process. Bulk RNA and single-cell RNA sequencing of the BMDM from the L13a-KO mice show widespread change in overall gene expression and robust differences in the diverse populations of the bone marrow-derived cells from the control and KO mice. In addition, this study also shows a substantial increase of Th1 and Th2 signature genes in naïve CD4+ T cells isolated from the L13a-KO animals. Together, our studies provide new insights into the regulations of macrophage polarization by L13a-driven novel intermediate effectors or mediators.
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: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: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.
