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(A) Microarray analysis of ARG gene expression. LNCaP cells cultured in 10% FBS were treated with 0, 1, 5 or 25 μM resveratrol for 48 h, and total RNA isolation and microarray analysis for ARG expression were performed as described in Materials and Methods. Triplicate treatments and analyses were conducted for each concentration. Results are expressed as a heat map. (Red color: upregulated gene and green color: downregulated gene.) Intensity of the color is proportional to the magnitude of effects on a log 2 scale, −3 to +3.
(B) RT–PCR analysis of effects of resveratrol on ARG expression. LNCaP cells cultured in 10% FBS were treated with or without resveratrol (25 μM) for 48 h, total RNA was isolated and mRNA levels of selected ARGs (B2M, PSA, SEPP1 and STK39) were determined as described in Materials and Methods. Results are expressed as mean ± SD (n = 3). Bar with * is significantly different from vehicle control at P < 0.05.
(C) RT–PCR analysis of effects of resveratrol on IGF-1R expression. LNCaP cells cultured in 10% FBS were treated with 0, 1, 5 or 25 μM resveratrol for 48 h, and total RNA was isolated and mRNA levels of IGF-1R determined as described in Materials and Methods. Results are expressed as mean ± SD (n = 3). Bars with *are significantly different from vehicle control (0) at P < 0.05.
(D) RT–PCR analysis of effects of resveratrol on CDKN1A expression. LNCaP cells cultured in 10% FBS were treated with 0, 1, 5 or 25 μM resveratrol for 48 h, and total RNA was isolated and mRNA levels of CDKN1A determined as described in Materials and Methods. Results are expressed as mean ± SD (n = 3). Bar with *is significantly different from vehicle control (0) at P < 0.05.
(E) Affinity of resveratrol for AR. AR binding assays were performed as described in Materials and Methods. Dihydrotestosterone (DHT) was used as positive control. Results are expressed as percent control ± SD (n = 3). Points with *are significantly different from vehicle control (0) at P < 0.05.
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