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. Author manuscript; available in PMC: 2023 Oct 10.
Published in final edited form as: Cancer Cell. 2022 Sep 29;40(10):1190–1206.e9. doi: 10.1016/j.ccell.2022.09.007

Figure 2. Transcriptional silencing of MHC-II is mediated by the CtBP co-repressor complex.

Figure 2

(A)Targeted epigenetic CRISPR screen results. For IFN-γ+ sorts, cells were pulsed with IFN-γ 25 ng/mL for 48 hours prior to each sort. Bubble plots show the top 1,000 enriched genes identified, with components of the CtBP complex indicated in red. P values were calculated using the RSA algorithm (König et al., 2007).

(B)Schematic representation of the CtBP complex.

(C)Cell surface MHC-II expression in MV4-11 Cas9 cells transduced with a pool of 2 sgRNAs targeting the indicated genes or a control sgRNA. Bars depict the mean percentage of MHC-II+ cells from 3 independent experiments indicated by points. Statistical analysis by unpaired t test compared to control cells; p value * < 0.05, ** < 0.01, *** < 0.001.

(D)Chromatin immunoprecipitation sequencing (ChIP-seq) plots for WIZ in MV4-11 cells at indicated genes.

(E)Cell surface MHC-II expression in MV4-11 cells treated with EHMT1/2 inhibitor (A-366) at the indicated doses or vehicle control in the presence or absence of IFN-γ 10 ng/mL for 24 hours. Representative plots from 2 experiments.

(F)CUT&Tag chromatin profiling of H3K9me2 and H3K27me3 at MHC-II genes in MV4-11 cells treated with A-366 1μM or vehicle control (ETOH) for 7 days. The HoxC cluster is included as a positive H3K27me3 control.

(G)Cell surface MHC-II expression in MV4-11 RREB1 KO cells transduced with a pool of 2 sgRNAs targeting STAT1 or a control sgRNA, and MV4-11 STAT1 KO or control cells treated with A-366 1μM for 7 days. Representative plots from 2 experiments.

See also Figure S2.