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. Author manuscript; available in PMC: 2020 Dec 21.
Published in final edited form as: Oncogene. 2020 Jun 21;39(31):5390–5404. doi: 10.1038/s41388-020-1370-9

Figure 2: Androgen indirectly induces Bnip3 through integrin α6β1 and HIF1α.

Figure 2:

A) Levels of PSA and Bnip3 mRNA in laminin-adherent C4–2 cells over a 24 h time course following treatment with 10nM R1881 measured by qRT-PCR and normalized to time 0. *p<0.05 relative to time 0, n=3 biological replicates, error bars = SD. B) Levels of Bnip3 mRNA in laminin-adherent VCaP, LNCaP, and C4–2 cells treated with (+) or without (-) 10nM R1881 in the absence (Veh) or presence of 10μg/mL cycloheximide (CHX) for 24 hours. Expression is relative to vehicle control. *p<0.05 relative to vehicle, n=3 biological replicates, error bars = SD. C) Levels of integrin α6 (ITGα6), Bnip3, PSA, and GAPDH in the cytosol (Cyto) and levels of androgen receptor (AR), HIF1α, and histone 3 (HH3) in the nucleus (Nuc) of laminin-adherent C4–2 cells treated with 10nM R1881 over a time course of 24 h as measured by immunoblotting. D) Laminin-adherent C4–2 cells stably expressing Tet-inducible shRNA targeting integrin α6 (Tet-shITGα6) were treated with (+) or without (-) doxycycline (Dox) for 48 h, and then stimulated with (+) or without (-) 10nM R1881 for 24 h. The levels of integrin α6 (ITGα6), Bnip3, PSA, and GAPDH in the cytosol (Cyto) and androgen receptor (AR), HIF1α, and histone 3 (HH3) in the nucleus (Nuc) were assessed by immunoblotting. E) Laminin-adherent C4–2 cells were transiently transfected with HIF1α or scrambled siRNA and 48 h later treated with 10nM R1881 for 24 h. The levels of Bnip3, PSA, and GAPDH in the cytosol (Cyto) and AR, HIF1α, and histone H3 (HH3) in the nucleus (Nuc) were assessed by immunoblotting.