Fig. 5

VCP inhibition enhances αS seeding in neurons. A Immunofluorescence for phospho-αS and Tuj1 (neurite marker) in HNs treated for 4 hours with αS PFF or monomer (1μg/ml) followed by washout and harvested 5 days later. B Immunofluorescence for phospho-αS and Tuj1 in HNs co-treated with αS PFF or monomer (1μg/ml) and LLoMe (1µM) or vehicle for 4 hours. Immunostaining wasperformed after 5 days. Scale bar =20μm. C Quantitation of phospho-αS/Tuj1 staining in 5B. Quantitation in 5B and subsequent HN studies were performed using the average intensity of multiple fields from individual coverslips. Each coverslip was treated as an independent experiment. n=6 for ethanol and LLoMe groups respectively. Experiments were repeated from 3 different cultures. **p< 0.01 by student’s t-test. Data are mean±S.E.M. D Immunofluorescence for phospho-αS and Tuj1 in HNs co-treated with αS PFF or monomer (1μg/ml) and ML240 (100nM) or DMSO control for 4 hours and harvested after 5 days. Scale bar =20μm. E Quantitation of phospho-αS/Tuj1 staining in 5D. n=6 for DMSO and ML240 group. Experiments were repeated from 3 different cultures. *p< 0.05 by student’s t-test. Error bars are ±S.E.M.)