(C) ASC J9 blocked testosterone-induced cell loss in an OS environment, but it did not influence H2O2-induced cell loss. (58C64). Depending on the level of OS in the Neochlorogenic acid cell, testosterone-induced OS could be neuroprotective via a preconditioning mechanism (58, 65, 66) or damaging by exacerbating existing OS damage (58C60, 64, Neochlorogenic acid 67C69). Using cell-impermeable androgens (value 0.05 indicates statistically significant differences. Comparisons were made by two- or three-way ANOVA using inhibitors, oxidative stressor, and hormone as impartial factors. Fisher least significant difference analysis was used to assess differences between the various groups. Each experiment was replicated at least three times with different cell cultures. Results mAR interacts with NOX1, NOX2, and G 0.05). In the presence of OS, testosterone further decreased cell viability, as indicated by a significant conversation between OS and hormone ( 0.05). The AR antagonist bicalutamide had no effect on cell viability, regardless of the OS environment. Furthermore, bicalutamide did not block testosterones unfavorable actions on cell viability in an OS environment. Open in a separate window Physique 3. Testosterones detrimental effects are not mediated through the classical genomic pathway. Testosterone alone does not affect cell viability. H2O2 induced Itga1 20% cell loss, which Neochlorogenic acid was exacerbated by testosterone. (A) The AR antagonist bicalutamide did not block testosterones negative effects in an OS environment. (B) Testosterone did not alter AR45 expression. The AR degrader, ASC J9, significantly decreased the Neochlorogenic acid expression of AR45, irrespective of Neochlorogenic acid the presence of testosterone. (C) ASC J9 blocked testosterone-induced cell loss in an OS environment, but it did not influence H2O2-induced cell loss. Results are reported as mean SEM. Results were determined by ANOVA followed by a Fisher least significant difference test. * 0.05 vs all groups; # 0.05 vs control; + 0.05 vs HT groups. B, bicalutamide; C, vehicle control; H, H2O2; HT, posttreatment T; J9, ASC J9; T, 100 nM testosterone. Because classical AR antagonists did not influence testosterones effects, an AR degrader, ASC J9, was used to degrade both cytosolic and mARs. ASC J9 selectively promotes AR degradation by disrupting the conversation between AR and AR coregulators, without impacting AR mRNA expression (86). To ensure that ASC J9 degraded mAR protein (45 kDa) expression, ASC J9 was used with and without testosterone, as previous reports stated that testosterone can stabilize AR (70, 87, 88). Consistent with our prior findings (70), testosterone did not alter mAR protein expression (Fig. 3B). In contrast, ASC J9 significantly reduced mAR expression ( 0.05), regardless of testosterone exposure (Fig. 3B). To determine whether degrading the mAR impacts testosterones negative effects in an OS environment, N27 cells were pretreated with ASC J9 for 2 hours prior to exposure to H2O2 and testosterone (Fig. 3C). As expected, significant negative effects from OS exposure on cell viability were observed ( 0.05). Additionally, significant interactions between oxidative stressor and testosterone treatment ( 0.05) and between oxidative stressor, hormone, and degrader ( 0.05) were observed. Specifically, H2O2 induced 20% cell loss, and testosterone exacerbated H2O2-induced cell loss. The AR degrader blocked testosterones negative effects on cell viability in an OS environment. However, the AR degrader did not impact H2O2-induced cell loss. The role of G protein and InsP3R in testosterone-induced neurodegeneration Because AR45 complexes with G 0.05), hormone ( 0.05), and an conversation between oxidative stressor and hormone ( 0.05) were observed. As expected, we observed no effects of testosterone alone and significant effects of H2O2 around the cell viability 20% cell loss. Testosterone, in the presence of an oxidative stressor, caused a further decrease in cell viability. However, GDPtest. * 0.05 vs all groups; # 0.05 vs control; + 0.05 vs HT groups. C, vehicle control; G, GDP 0.05), hormone ( 0.05), and an conversation between oxidative stressor and hormone ( 0.05) (Fig. 4B). Again, no effects of testosterone alone were observed. H2O2 significantly induced 20% cell loss and testosterone worsened H2O2s damaging effects. Similar to the GDP analog, the G 0.05), hormone ( 0.05), inhibitor ( 0.05), an conversation between oxidative stressor and hormone ( 0.05), and an conversation between oxidative stressor, hormone, and InsP3R.