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Fig. 1 | Genome Biology

Fig. 1

From: MBD5 and MBD6 stabilize the BAP1 complex and promote BAP1-dependent cancer

Fig. 1

Endogenous MBD5 and MBD6 are stable components in the BAP1 complex. A HEK293T cells were infected by lentivirus expressing GFP, GFP-tagged MBD5, or GFP-tagged-MBD6. The protein levels of GFP, GFP-MBD5, and GFP-MBD6 were determined by western blot, n=2. B The GFP-fusion proteins were purified from HEK293T cells stably expressing GFP, GFP-MBD5, or GFP-MBD6 defined in A. The purified proteins were subjected to mass spectrometry analysis. Peptide numbers of subunits within the BAP1 complex pulled down by GFP-tagged proteins are shown. C Immunoprecipitation (IP) of endogenous BAP1 from HEK293T cells followed by immunoblot (IB) for BAP1, MBD5, and MBD6. IgG was used as a negative control, n = 2. D IP of endogenous MBD5 or MBD6 from HEK293T cells followed by IB for MBD5, MBD6, BAP1, ASXL1, and ASXL2. IgG was used as a negative control, n = 2. E Nuclear extract from HEK293T cells was subjected to size exclusion (SE) chromatography and then protein levels of BAP1, MBD5, and MBD6 were determined by western blot analysis, n = 2. F The average plots shown represent the chromatin occupancy of H3K27Ac, H3K4me1, H3K4me3, and H3K27me3 centered on MBD5 peaks (F) or MBD6 peaks (G). H Sorted and centered heatmaps generated from ChIP-seq data analyses show the occupancy of BAP1, MBD5, and MBD6 in HEK293T cells. All rows are centered on BAP1 peaks based on the ranking of signals. I The average plot shows the co-occupancy between BAP1, MBD5, and MBD6 centered on BAP1 peaks. J The representative tracks show H3K27Ac, H3K4me1, and H3K4me3 levels at BAP1, MBD5, and MBD6 occupied loci in HEK293T cells. K A Venn diagram representation of the overlapped ChIP-seq peaks between BAP1, MBD5, and MBD6

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