Fig. 4

VCP is required for autophagy-mediated Tau clearance. a Western blots of VCP, total Tau (T46) and pTau (AT8) of vcp MO and mRNA-injected embryos. vcp MO knockdown resulted in accumulation of Tau proteins, which was rescued by human VCPWT overexpression but not by mutant human VCPDKO. b Representative confocal images and the quantification of primary motor axons of vcp morphants and VCP mRNA expression in (a) at 2 dpf, visualized by Znp1 immunostaining (left panels) and tracing of motor axons for quantitation using images J (right panels). vcp knockdown showed more than ~ 50% reduction in both length and branching numbers of primary motor axons, which were partially rescued by VCPWT expression but not by mutant VCPDKO. c The diagram shows mutations in VCP protein construct; western blots of VCP, total Tau (T46), pTau (AT8) and autophagy marker (LC3, p62) antibodies of vcp MO and mRNA-injected embryos. vcp MO knockdown resulted in accumulation of Tau proteins, which was rescued by human VCPWT and VCPD395G overexpression but not by VCPDKO, VCPA232E, or VCPR155H. d Representative confocal images and the quantification of primary motor axons of vcp morphants and VCP mRNA expression in (c) at 2 dpf, visualized by Znp1 immunostaining (left panels) and tracing of motor axons for quantitation using images J (right panels). vcp knockdown showed more than ~ 50% reduction in both length and branching numbers of primary motor axons, which were partially rescued by VCPWT and VCPD395G expression but not by mutant VCPDKO, VCPA232E, or VCPR155H. Data represent mean ± SEM, n = 10 (10 individual animals), ANOVA test, *p < 0.05; **p < 0.01; ***p < 0.005; Scale bar = 200 µm