Fig. 6

Maged1 deficiency attenuates impairment of autophagy in MPP⁺-intoxicated cells and MPTP-treated mice. A SH-SY5Y cells were transfected with GFP-LC3 and visualized using confocal microscopy. Scale bar:10 μm. B Quantification of GFP-LC3 puncta formation in (A), n = 6 different fields. C SH-SY5Y cells were transfected with NC or si-Maged1 and treated with 200 μM MPP⁺ for 48 h and cell lysates were detected for LC3 and P62. D, E Quantification of LC3II/GAPDH (D) and P62/GAPDH (E) from the results of (C). F Three days after the final administration of MPTP, mice midbrains were collected and protein expression of LC3 and P62 was measured using western blot; GAPDH was used as a loading control. G, H Quantification of LC3II/GAPDH (G) and P62/GAPDH (H) from results of (F), n = 4 for each group. I, J Western blot analysis was used to detect protein levels of p-Akt (S473), p-Akt (T308), t-Akt, p-mTOR (S2448), t-mTOR, p-p70S6K (T389), t-p70S6K, p-S6 (S235/236), t-S6, and Maged1 in NC or si-Maged1-transfected SH-SY5Y cells. For A-I, experiments were performed in triplicate. Data are shown as means ± SE. * P < 0.05, ** P < 0.01, *** P < 0.001, ^#P < 0.05, ^##P < 0.01, ^###P < 0.001. NC normal control, KD knock down