Figure 3. Adaptive de novo expression of the 20S proteasome diminishes with age in a sex-dependent manner. (A,B) Virgin females of the Actin-GS-255B strain were crossed to males of the w strain and progeny were assayed for 20Sα expression without H2O2 pretreatment, or after pretreatment with [10µM or 100µM] H2O2. (A) 3 day and 60 day aged females. (B) 3 day and 60 day aged males. All samples were compared to the 3 day old 0µM H2O2 controls (C) Basal expression of the 20S proteasome α subunits was measured between 3 day old females, 60 day old females, 3 day old males, and 60 day old males, with samples normalized to the 3 day old female. Western blots were performed in triplicate, normalized to Actin-HRP, and quantified using ImageJ. Error bars denote standard error of the mean (S.E.M) values. * P <0.05 and ** P < 0.01, relative to control using one-way ANOVA. Asterisks color indicates the age of the sample, young female (pink *) and aged female (black *). All statistical significance was calculated relative to the young controls.