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. Author manuscript; available in PMC: 2016 Mar 29.
Published in final edited form as: Science. 2016 Jan 21;351(6274):aad5510. doi: 10.1126/science.aad5510

Fig. 5. MafB inhibits macrophage self-renewal by direct repression of self-renewal gene enhancers.

Fig. 5

A) Colony assays for Maf-DKO macrophages expressing empty vector (-MafB) or a doxycycline-inducible flag-tagged MafB allele (+MafB) counted after 14 days of culture in methocult medium containing M-CSF and doxycycline (DOX), showing culture dishes (0.63x), and number of colony-forming units (CFU). Error bars represent the SD of two technical replicates and data are representative of three independent experiments. B) Expression of self-renewal genes in Maf-DKO macrophages (-MafB) and Maf-DKO macrophages expressing a doxycycline-inducible, flag-tagged MafB allele (+MafB) after 2 hours stimulation with M-CSF determined by nano-fluidic real-time PCR on Fluidigm array. Data are representative of three independent experiments. C) Aggregation plots showing average ChIP-seq signals for P300 and H3K27ac in Maf-DKO, WT BMM and Maf-DKO macrophages expressing a doxycycline-inducible, flag-tagged MafB allele in the presence of doxycycline (Maf-DKO+MafB) for the self-renewal associated enhancers regions (total=88 regions). D) Direct alignment of regions proximal to Maf-DKO-only enhancers for flag-MafB binding in Maf-DKO+MafB and corresponding regions in Maf-DKO and Maf-DKO+MafB macrophages for P300 and H3K27ac binding. E) Histogram showing the percent of WT BMM-only, Maf-DKO-only and self-renewal gene-associated enhancers bound by MafB as determined by ChIP-seq for flag-MafB in Maf-DKO+MafB cells. F) Genomic regions surrounding MYC gene with Chip-seq tracks as labelled.