Fig. 3: LSD1 requires the AR AF-2 surface and its catalytic activity to transactivate AR.

a Scheme of AR and LSD1 modular domains and specific motifs. Numbers refer to AR NM_000044 and LSD1 NM_001009999. NTD, amino-terminal domain; DBD, DNA-binding domain; LBD, ligand-binding domain; AOD, amine oxidase domain; CTD, carboxy-terminal domain. b Transcriptional assay in HEK293T cells expressing AR55Q or the AF-2 mutant AR55Q-E897K, alone (mock) or with LSD1. Cells were treated with DHT (10 nM, 16 h; n = 3 biological replicates). c (Left) Transcriptional assay in HEK293T cells expressing AR65Q alone (mock) or with either LSD1 or LSD1-LXXAA. Cells were treated with DHT (10 nM, 16 h; n = 6 biological replicates mock, n = 3 biological replicates LSD1 and LSD1-LXXAA). (Right) Western blot of LSD1 and LSD1-LXXAA expression in HEK293T cells. Shown is one representative image of three biological replicates. d Transcriptional assay in HEK293T cells expressing AR65Q alone (mock) or with either LSD1 or the catalytic inactive mutant LSD1-K685A. Cells were treated with DHT (10 nM, 16 h; n = 3 biological replicates). e Transcriptional assay in HEK293T cells expressing AR65Q treated with DHT only or together with SP-2509 (100 nM) or TCP (10 μM) for 16 h (n = 3 biological replicates). f Western blot of H3K4me2 in C2C12 cells differentiated to myotubes for 10 days in the presence of DHT (10 nM). H3K4me2 was detected with a specific antibody that recognize the H3 modified K residue. H3 antibody was used as loading control (n = 4 biological replicates). Graphs show mean ± SEM; two-way (b), one-way (c, d, e) ANOVA followed by Tukey HSD tests, or two-tailed Student t-test (f). Source data are provided as a Source data file.