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. Author manuscript; available in PMC: 2019 Jul 15.
Published in final edited form as: Cancer Res. 2018 May 15;78(14):4022–4035. doi: 10.1158/0008-5472.CAN-17-3728

Figure 1.

Figure 1

C86 is an inhibitor of the AR/ARv7 signaling axis in CRPC cells.

(A) RT-qPCR of AR/ARv7 target genes, normalized to 18S rRNA, from 22Rv1 CRPC cells treated with 5 μM of each compound or bicalutamide (Bic) for 20 hours. Data represent a mean of three replicates +/− S.D. PSA (prostate specific antigen) refers to gene designation KLK3.

(B) WB of cell lysate from 22Rv1 CRPC cells treated with increasing concentrations of C86 (left) or 10 μM C86 for up to 24 hours (right).

(C) WB of cell lysate from VCaP CRPC cells treated with increasing concentrations of C86 for 6 hours.

(D) WB of soluble AR, ARv, and ARv7 from 22Rv1 CRPC cells following pre-incubation with 10 μM MG132 and/or 10 μg/mL cycloheximide (CHX) for 1 hour, followed by +/− 10 μM C86 for 6 additional hours. Equal amounts of protein (10 μg) were loaded for WB of soluble and insoluble fractions (see also Supplemental Figure 1D).

(E) Viability of 22Rv1 CRPC cells exposed to increasing concentrations of C86 or anti-androgens (MDV3100 - enzalutamide, Bic) for 72 hours as determined by MTT assay. Data represent a mean of four to six replicates +/− S.D.

Experiments were repeated at least three times. AR antibody N-20 recognizes full-length AR (FL-AR) and likely also a number of similarly sized splice variants.