Figure 5.

PI3-kinase pathway activation in DHT-mediated VEGF production. A, Level of VEGF in fibroblast CM measured by ELISA after 48 hours treatment with 40nM DHT in the presence or absence of 10μM PI3-kinase inhibitor (LY294002). Statistical analysis was performed using one-way ANOVA with Bonferroni correction. *, P < .01 vs untreated control; n = 3 donors performed in triplicate over 3 individual experiments (means ± SEM). B, The level of pAKT(Ser473)/AKT expressed in fibroblasts from young men after 48 hours treatment with 40nM DHT in the presence or absence of 4μM HF or 10μM PI3-kinase inhibitor (LY294002). Data are normalized to vehicle control; n = 3. Statistical analysis was performed using one-way ANOVA with Bonferroni correction. *, P < .05, DHT vs untreated control; #, P < .01, DHT plus HF vs untreated control plus HF. C, The level pAKT(Tyr308)/AKT expressed in fibroblasts from young men treated for 48 hours with DHT with or without the addition of LY294002. D, The level of pAKT(Ser473)/AKT expressed in young fibroblasts transfected with scrambled siRNA or AR siRNA after 48 hours treatment with 40nM DHT; n = 3. Data are normalized to untreated cells transfected with scrambled siRNA or AR siRNA. Statistical analysis was performed using one-way ANOVA with Bonferroni correction. *, P < .05, DHT vs control scrambled siRNA transfected cells. E, The level of pAKT(Ser473)/AKT expressed in fibroblasts from young and old men after 48 hours treatment with 40nM DHT. Statistical analysis was performed using 2-way ANOVA with Bonferroni correction. *, P < .05 vs young control; n = 3 donors performed in duplicate over 3 individual experiments (means ± SEM).