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. 2018 Jan 8;115(4):798–803. doi: 10.1073/pnas.1700465115

Fig. 5.

Fig. 5.

PFF-induced pathology in GBA1-PD hDA neurons is reduced by miglustat treatment. (A) Fluorescent analysis of p-α-syn levels and TH staining in 5 μg/mL PFF-treated hDA neurons for 10 d without or with 100 μM miglustat treatment for 3 d before adding PFF. The images were generated by the tile scan algorithm in the Zen software (Carl Zeiss). [Scale bars, 100 μm for low magnification (Upper, 40×, 8 × 8 tile image) and 50 μm for high magnification (Lower, cropped image from low-magnification image.] (B) Quantification of p-α-syn levels for A (n = 6). (C and D) Control and GBA1-PD hDA neurons were sequentially fractionated in 1% Triton X-100 (TX-soluble) followed by 2% SDS (TX-insoluble) 14 d after 5 μg/mL PFF treatment. Western blots from (C) TX-soluble and (D) TX-insoluble fractions with or without 100 μM miglustat-treated hDA neurons using anti–α-Syn, anti-TH, anti–p-α-syn, and anti–β-actin antibodies. (E and F) Cell death in 5 μg/mL PFF-treated hDA neurons for 14 d without or with 100 μM miglustat treatment for 3 d before adding PFF was assessed using (E) AlamarBlue (n = 6) and (F) LDH cytotoxicity assay (n = 5). The error bars represent SEM. *P < 0.05; **P < 0.01; ***P < 0.001; n.s., not significant.