MANF Inhibits α-Synuclein Accumulation through Activation of Autophagic Pathways

Figure 7

MANF inhibits the accumulation of SNCA^A53T in PD cellular model by activating macroautophagy. (a)–(d) Effects of MANF on the levels of SNCA^A53T and macroautophagic-related protein expression. SNCA^A53T SH-SY5Y cells were treated with MANF (250 and 500 ng/ml) and Dox (600 ng/ml) for 24 h (a, b) or 48 h (c, d), respectively; then, the protein levels of SNCA^A53T, LC3-I/II, Beclin-1, and P62 were detected by western blot analysis. (e, f) SNCA^A53T SH-SY5Y cells were treated with autophagy inhibition CQ (10 μM) for 24 h, and cell lysates were immunoblotted by anti-LC3 and anti-Beclin-1 antibodies. (g, h) SNCA^A53T SH-SY5Y cells were treated with MANF (500 ng/ml), Dox (600 ng/ml), and CQ (10 μM) for 48 h, and the protein level of SNCA^A53T was detected by western blot analysis. Data were expressed as from three independent experiments. ^#, ^##, and ^$ vs. control group; vs. Dox-treated group; ^& vs. combined treatment with MANF and Dox group.

(a)

(b)

(c)

(d)

(e)

(f)

(g)

(h)