Fig. 4: LSD1 and PRMT6 synergistically transactivate normal and polyQ-expanded AR.

a Proximity ligation assays analysis in MN1 cells expressing AR24Q or AR100Q and vectors for Lsd1 and Prmt6 silencing. Cells were treated with DHT (10 nM, 16 h). Scale bar = 17 µm. Graphs show quantification of nuclei from three independent experiments (AR24Q cells: P6/LSD1 vehicle n = 12, DHT n = 22; AR/LSD1 Cas9 scrambled n = 169, Cas9 shPrmt6 n = 198; AR/PRMT6 Cas9 scrambled n = 184, Cas9 g2 n = 237. AR100Q cells: P6/LSD1 vehicle n = 79, DHT n = 116; AR/LSD1 Cas9 scrambled n = 302, Cas9 shPrmt6 n = 190; AR/PRMT6 Cas9 scrambled n = 202, Cas9 g2 n = 164. b Western blots of MN1 cells transduced with lentiviral vectors for Lsd1 and Prmt6 silencing (n = 3 biological replicates). LSD1 g2 **p = 0.0036, g2/sh#1 *p = 0.048, g2/sh#2 **p = 0.0071. PRMT6 sh#1 ***p = 0.00001, sh#2 ***p = 0.0000004, g2/sh#1 *p = 0.011, g2/sh#2 **p = 0.003. c Immunoprecipitation of PRMT6 and immunoblotting of the indicated proteins in the skeletal muscle (quadriceps) of 24-week-old WT and knock-in AR113Q mice (n = 3 mice/genotype). d Transcriptional assays in HEK293T cells expressing AR24Q or AR65Q alone (mock) or with LSD1 and PRMT6 treated for 16 h with vehicle, DHT (10 nM), TCP (10 μM), or Adox (10 μM) (AR24Q: n = 7 vehicle, n = 3 TCP, Adox, TCP + Adox. AR65Q: n = 7 vehicle, n = 4 TCP, Adox, TCP + Adox). e (Left) Transcriptional assay in HEK293T cells expressing AR24Q or AR65Q with or without CRISPR guides to silence Lsd1 and Prmt6 (n = 3 biological replicates). (Right) Western blot of PRMT6 levels in HEK293T expressing Cas9 alone or together with guides to silence PRMT6 (n = 3 biological replicates). Cas9/g1 ***p = 0.00048, Cas9/g2 ***p = 0.00056.AR, LSD1, and PRMT6 were detected with specific antibodies, and β-Tub and CNX were used as loading controls. Graphs show mean ± SEM; two-tailed Student t-test (a, b) or one-way ANOVA followed by Tukey HSD tests (d, e). Source data are provided as a Source data file.