Fig. 4

Effects of Cu on Cho release. A ABCA1 levels determined by western blot (n = 8); B Labeled Cho (¹⁴Cho) produced by de novo synthesis determined by HP-TLC followed by the analysis of intensity of radioactive bands (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 significant differences respect to the control of the same sex, ^# significant differences between Cu female vs Cu male (p < 0.05 for ABCA1 and p < 0.01 for ¹⁴Cho release), and ^&p < 0.01 significant differences between Cu and THB treatments in the same sex