Fig. 3
From: Coordinate regulation of mutant NPC1 degradation by selective ER autophagy and MARCH6-dependent ERAD
Accumulating I1061T NPC1 does not traffic to lysosomes. a Primary human fibroblasts expressing I1061T or P1007A/T1036M NPC1 were treated for 24 h with vehicle, 10 μM MG132 (MG), 100 nM bafilomycin A1 (Baf), or 1 mM cyclodextrin (Cyclo). Unesterified cholesterol was labeled with filipin (blue), and staining intensity calculated from three independent experiments (16 fields/experiment). Scale bar = 50 μm. Dashed lines outline plasma membrane. Quantified at the bottom. b CTRL, I1061T, or P1007A/T1036M NPC1 fibroblasts were treated for 24 h with vehicle (Veh), 100 nM bafilomycin A1 (Baf), or 100 nM epoxomicin (Epox). Lysates were digested with endoglycosidase H (E), PNGaseF (P), or not treated (NT) and then analyzed by western blot. Data are mean ± s.e.m. n.s., not significant, *P ≤ 0.05, ****P ≤ 0.0001 by ANOVA with Tukey’s posthoc test F = 50.85
