Supplementary Figure 5: KDM1A K114me2 regulates AR-dependent gene expression by controlling chromatin occupancy of CHD1 and AR. | Nature Structural & Molecular Biology

Supplementary Figure 5: KDM1A K114me2 regulates AR-dependent gene expression by controlling chromatin occupancy of CHD1 and AR.

From: Assembly of methylated KDM1A and CHD1 drives androgen receptor–dependent transcription and translocation

Supplementary Figure 5

(a-f) ChIP analyzes performed with anti-KDM1A, anti-KDM1A K114A, anti-EHMT2, anti-CHD1, anti-AR antibodies and rIgG. Samples originate from LNCaP cells that were cultivated in the presence or absence of DHT, treated with or without Bix-01294 (a-c) or transfected with siRNA (d-f) as indicated. The precipitated chromatin was quantified by qPCR analysis using primers flanking AREs in the enhancer regions of the KLK3 ( a,b,e,f) or TMPRSS2 (d) genes and an unrelated control region (c) as indicated. (g-i) Expression analyzes of androgen-regulated genes. Samples originate from LNCaP (g,i) or LAPC4 (h) cells treated with Bix-01294 (g,h) or RNAi against EHMT2, KDM1A, or CHD1 (i). (j) Expression analyzes. Samples originate from LNCaP cells cultured in the absence or presence of DHT and treated with RNAi against KDM1A or CHD1. (d,f,i) Anti-tubulin (d,f,i), anti-KDM1A (d), anti-EHMT2 (f) and anti-CHD1 (i) Western blots. Samples are lysates of LNCaP cells transfected with an unrelated siRNA control (Ctrl) or a siRNA against EHMT2 (f), CHD1 (i), or KDM1A (d). Error bars, s.d. (a-f) Technical replicates of one representative experiment out of three (n=3). (g-j) Biological replicates (n=3 cell cultures). *** p<0.0001; ** p<0.001; * p<0.005 by two-tailed Student’s test.

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