Fig. 1: HMMR is suppressed by AR knockdown but not by treatment with ARSIs.

a Volcano plot of differentially expressed genes affected by either siAR versus siCON (red; n = 599 genes), or Veh versus ENZ (black; n = 23 genes). b Validation of HMMR expression in response to siAR or 1 µM ENZ by RT-qPCR. Gene expression was normalised to GUSB and L19. Data is presented as mean ± SD of 3 biological replicates and are representative of 3 independent experiments. Controls (Veh and siCON) were set to 1 and data statistically evaluated using two-way ANOVA with Tukey’s multiple comparison test (treatment vs control; ****p < 0.0001). c AR and HMMR protein expression in response to siAR or 1 µM ENZ by Western Blot. GAPDH was used as a loading control.KLK3, KLK2, FKBP5 and TMPRSS2 expression in response to treatment with increasing dose of ARSIs apalutamide (d) or darolutamide (e) for 24 h, by RT-qPCR. Gene expression was normalised to GUSB and L19. Data are presented as mean ± SD of 3 biological replicates and are representative of three independent experiments. Control (Veh) was set to 1 and data was statistically evaluated using one-way ANOVA with Dunnett’s multiple comparison test. (*p < 0.05, **p < 0.01, ****p < 0.0001). AR, PSA and HMMR protein expression in response to treatment with ARSIs apalutamide (f) or darolutamide (g) for 24 h, by Western Blot. Numerals above each lane represent densitometric analysis of each protein relative to loading controls GAPDH (f) or β-Actin (g).