Fig. 2
Cu treatment effect on oxidative stress in female and male astrocytes and cellular Cu content. A Cellular Cu concentration (µg/dL) measured by spectrophotometry (n = 6); B ROS levels were determined using DCF-DA by flow cytometry (n = 6); C SOD activity was measured by spectrophotometry (n = 6 to 8); and D Lipid peroxidation was determined by the TBARS assay (n = 8). All determinations were made after 24 h of Cu treatment (400 µM of CuSO4; gray bar). Untreated cells were used as controls (black bar), TBH (500 µM) was used as a positive control of ROS production (dark gray bars), and NAC (10 µg/mL) was used as an antioxidant molecule to counteract Cu ROS generation to be used as a second negative control (light gray bars). Results were calculated using 2-way ANOVA plus Bonferroni’s multiple comparisons test, and data expressed mean ± SD, **p < 0.01 and ***p < 0.001 significant differences respect to the control of the same sex, #significant differences between female and male astrocytes with the same treatment (p < 0.05 for TBH,  p < 0.01 for Cu ROS and p < 0.1 for Cu SOD activity), and + p < 0.05 significant differences between Cu and Cu + NAC treatment of the same sex