Figure 5.

STAT3 regulates miR-204/TRPLM1 activity to mediate mitochondrial damage, ROS production, and mitochondrial autophagy

(A) Expression of STAT3/p-STAT3 normalized to β-actin in AD-modeled neurons detected by western blot analysis. (B) Expression of miR-204 detected by qRT-PCR is shown. (C) The expression of PINK1, Parkin, LC3II, and Beclin1 normalized to β-actin in AD neurons detected by western blot analysis is shown. (D) The mitochondrial membrane potential of AD-modeled neurons detected by JC-1 staining is shown, with green referring to JC-1 monomer and golden color referring to JC-1 polymer. (E) Cell ATP production is shown. (F) Total ROS production in AD-modeled neurons detected by DCFH-DA staining is shown. DCFH-DA without inherent fluorescence can enter into cells through the membrane and then be hydrolyzed into DCFH by esterase. DCFH is incapable of crossing through the membrane, so that probes can be easily loaded into cells where active oxygen can oxidize non-fluorescent DCFH to generate DCF with green fluorescence, which indicates the ROS content. (G) ROS production in mitochondria in AD-modeled neurons detected by DHR123 staining is shown. DHR123 was oxidized into rhodamine123 by ROS in the mitochondria, and rhodamine123 was excited at 488 nm while green fluorescence was emitted at 515 nm. ∗p < 0.05. Measurement data were expressed as mean ± standard deviation. The comparison between two groups was performed by independent sample t test. The experiment was repeated three times independently.