Figure 4.

Downregulation of miR-204 inhibits mitochondrial damage, ROS production, and mitochondrial autophagy induced by Aβ1-42 through upregulating TRPML1

(A) Expression of mitochondrial-autophagy-related proteins normalized to β-actin in AD-modeled neurons detected by western blot analysis. (B) The mitochondrial membrane potential of neurons detected by JC-1 staining is shown, where green refers to JC-1 monomer and golden color refers to JC-1 polymer. (C) Cell ATP production is shown. (D) Total ROS production in AD neurons detected by DCFH-DA staining is shown. DCFH-DA without intrinsic 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 ROS content. (E) 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. Measurement data were expressed in the form of mean ± standard deviation. The comparison between two groups was performed by independent sample t test. ∗p < 0.05. The experiment was repeated three times independently.