A) MC65 cells were treated with ZCM-I-1 in the presence of tetracycline (+TC) or absence of tetracycline (−TC) conditions for 48 h. After incubation with mitoSOX (2.5 μM), cells were analyzed by flow cytometry; MC65 cells (B) or mouse cortical neurons (DIV14) (C) were treated with vehicle or ZCM-I-1 (1 μM) for 12 h, then the cells were collected, washed, and suspended in HBSS. Upon addition of mitoSOX (2.5 μM) and rotenone (500 nM), the samples were analyzed by flow cytometry; SHSY5Y cells (D) or mouse cortical neurons (DIV14) (E) were treated with ZCM-I-1 at indicated concentrations or NAC (10 μm) or trolox (10 μM) and MPP+ (30 μM) for 24 h. After incubation with TMRM (100 nM), the samples were analyzed by flow cytometry; F) ROS production in permeabilized mouse brain mitochondria (N=6) pretreated with ZCM-I-1 was measured upon addition of NADH (10 μM) using Amplex red and HRP. Data presented as mean ± SEM. Statistical analysis by student t-test.