Figure 5: VEGF replenishment reverses defective autophagy in NP–C mice.
From: Pathological roles of the VEGF/SphK pathway in Niemann–Pick type C neurons
(a) Western blot analysis of LC3, beclin-1, p62 and cathepsin D in primary cultured PNs derived from WT, NP–C and VEGF/NP–C mice (WT, n=5; NP–C, n=6; and VEGF/NP–C, n=6). (b) Immunocytochemistry of LC3 in WT, NP–C and VEGF/NP–C PNs (n=6 per group; scale bar, 20 μm). (c) Cathepsin D activity in primary cultured PNs (WT, n=5; NP–C, n=6; and VEGF/NP–C, n=6). (d) Western blot analysis of LC3, beclin-1, p62 and cathepsin D in the cerebellums of 6-week-old WT, NP–C and VEGF/NP–C mice (WT, n=6; NP–C, n=7; and VEGF/NP–C, n=7). (e) Cathepsin D activity in the cerebellums of WT, NP–C and VEGF/NP–C mice (WT, n=5; NP–C, n=6; and VEGF/NP–C, n=6). (f) EM images and quantification data of the cerebellum (n=5 per group; low-magnification scale bar, 1 μm; high-magnification scale bar, 200 nm). Arrow indicates autophagic vacuole. (g) Western blot analysis of Rab5 and Rab7 levels in the cerebellum (n=6 per group). (h) Cerebellar sections were immunostained with anti-active caspase-3 and the number of active caspase-3-positive cells in PCL was quantified (n=5 per group; scale bar, 50 μm). a–g, one-way analysis of variance, Tukey’s post hoc test. h, Student’s t-test. *P<0.05, **P<0.01, ***P<0.005. All error bars indicate s.e.m.
