Fig. 3.

AR-modulatory miRs stabilise AR transcript and regulate apoptosis and WT- and variant-AR-driven proliferation, and MiR-346, -361-3p and -197-3p inhibitors show additive effects with AR silencing. a, b SRB proliferation assay analysis of LNCaP/ARsiRNA cells transfected with miR-346 (a) or miR-361-3p (b) mimic (7.5 nM – miR-346, 20 nM – miR-361-3p) or inhibitor (10 nM – miR-346, 20 nM – miR-361-3p) ± 1 µM Dox to induce AR siRNA (ARKO) for 6 days. c SRB proliferation assay analysis of DU145 cells transfected with miR-346 (i) or miR-361-3p (ii) inhibitor (5 and 20 nM) for 6 days. d–g SRB proliferation assay analysis of 22RV1 (di, fi), 22RV1-AR-FL-KO (AR variant-driven) (dii, fii), CWR-R1-AD1 (ei, gi) and CWR-R1-D567^es (AR-v567^es-driven) cells (eii, gii) transfected with: d, e miR-346 inhibitor (5 and 20 nM) or f, g miR-361-3p inhibitor (5 and 20 nM) for 6 days. a–g Data are presented relative to absorbance at day 0. Points: mean absorbance at 492 nm for three independent experiments performed in quadruplicate ± SEM. h, j qRT-PCR analysis of (h) AR and (j) PSA mRNA levels in LNCaP/ARsiRNA cells transfected with miR-346 (i) or miR-361-3p (ii) inhibitor or mimic (20 nM) ± Doxycycline (1 µM) for 24 h. L19 was used as a normalisation gene. Columns: mean ± SEM for three independent experiments performed in triplicate. k Caspase 3/7 Glo assay analysis of apoptosis in LNCaP/AR siRNA cells ± miR mimic or inhibitor (20 nM) ± Doxycycline (1 µM) for 72 h. Columns: mean relative luminescence for three independent experiments performed in triplicate ± SEM. *P ≤ 0.05, **P ≤ 0.005, ***P ≤ 0.0001. See also Fig. S6