Figure 5: ROS produced by up-regulated MAOA plays an essential role for androgen deprivation-induced autophagy activation, NED and anti-apoptosis.

(A) Immunoblotting of MAOA, LC3B-II and p62/SQSTM1 in shCtrl and shMAOA LNCaP cells treated as described in Fig. 3C for 48 and 96 hours. β-actin was used as loading control. Uncropped images are presented in Supplementary Figure S13A (right). (B) ROS production measured by OxiSelect In Vitro ROS/RNS Assay Kit in cells treated as described in (A). Data represent mean ± S.D. (n = 3); ***p < 0.001 by Student’s-t test. (C) Immunoblotting analysis of MAOA, β-tubulin III, LC3B-II and p62/SQSTM1 in cells treated as described in Fig. 1B in the presence or absence of 5 mM NAC. β-actin was used as loading control (upper panel). ROS production was measured as in (B). Data represent mean ± S.D. (n = 3); ***p < 0.001 by Student’s-t test (bottom panel). Uncropped images are presented in Supplementary Figure S13B. (D) Inhibition of ROS reduces neurite extension of LNCaP cells treated as described in (C) for 96 hours. Average neurite elongation was quantified as described in Fig. 1B ***p < 0.001 by Student’s-t test. (E) Inhibition of ROS decreases expression of NE markers in LNCaP cells treated as described in (C) for 96 hours. Data represent mean ± S.D. (n = 3); ***p < 0.001 by Student’s-t test. (F and G) Inhibition of ROS decreased cell viability (F) and increased caspase 3 activity (G) of LNCaP cells treated as described in (C). Data represent mean ± S.D. (n = 3); ***p < 0.001 by Student’s-t test.