MANF Inhibits α-Synuclein Accumulation through Activation of Autophagic Pathways

Figure 5

Effects of Nrf2 on the early activation of macroautophagy by MANF. (a, b) SNCA^WT SH-SY5Y cells were treated with Lamp-2A-RNAi 2 plasmid for 24 h, followed by incubation with MANF (500 ng/ml) and Dox (600 ng/ml) for another 24 h. The levels of LC3, Beclin-1, and P62 were detected by western blot analysis. (c, d) SNCA^WT SH-SY5Y cells were treated with MANF (500 ng/ml) and Dox (600 ng/ml) for another 24 h or 48 h, respectively; cell lysates were immunoblotted by anti-Nrf2 antibody. (e, f) SNCA^WT SH-SY5Y cells were treated with Lamp-2A-RNAi 2 plasmid for 24 h, followed by incubation with MANF (500 ng/ml), ML385 (5 μM), and Dox (600 ng/ml) for another 24 h. The levels of LAMP-2A, Nrf2, LC3, Beclin-1, and P62 were detected by western blot analysis. Data were expressed as from three independent experiments. ^# vs. control group; vs. Dox-treated group; ^& vs. combined treatment with MANF and Dox group.

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