Prostate cells express megalin and import SHBG-bound T. A, Gene expression of LRP2, AR, VDR, and CYP27B1 in a panel of prostate cell lines as shown by qRT-PCR shown as relative quantitation to HPRT1. Error bars are SEM. (A, inset) Western blot analysis for megalin in prostate cell line panel. B, Regulation of KLK3 (PSA gene) expression after 24 hours following treatment with vehicle control (CTL), 50 nmol/L T alone (T), 50 nmol/L T preincubated with 500 nmol/L SHBG (SHBG), or T + SHBG in cells preincubated with 1 μmol/L MEG-Inh in LNCaP and 22Rv1 prostate cancer lines. C, Visualization (× 63) of SHBG-555 (red) import into cells with DAPI (blue) nuclear and F-actin (green) cytoskeletal counterstains. Statistical analysis was performed using a one-way ANOVA with a two-stage linear step-up procedure of Benjamini, Krieger, and Yekutieli for multiple comparisons; *, P < 0.05 for comparison with CTL, all graphs represent mean ± SEM of three or more individual experiments with two replicates per experiment.