Figure 3: NPC1 knockdown reduces VEGF expression and SphK activity.
From: Pathological roles of the VEGF/SphK pathway in Niemann–Pick type C neurons
(a–c) Primary cultures of normal and VEGFtg PNs were transfected with control or NPC1 siRNA. Three days after transfection, we measured the levels of Np c1 (a) and Vegf (b) mRNA and secreted VEGF protein (c) in PNs (n=7 per group). (d,e) SphK activity (d), sphingosine and S1P (e) were estimated in PNs transfected with control (n=6) or NPC1 siRNA (n=7). (f–j) Four-week-old WT and VEGFtg mice were injected with control or NPC1 shRNA into the cerebellum. Mice were sacrificed at 3 days after the injection. Np c1 (f) and Vegf (g) mRNA levels were estimated in LCM-captured PNs and VEGF protein levels (h) were measured in the cerebellums (n=7 per group). (i) Left, SphK activities were measured in the cerebellums (n=7 per group). Right, relative levels of Sphk1 mRNA from LCM-captured PNs samples (n=8 per group). (j) Sphingosine and S1P were measured in the cerebellums (n=7 per group). a–j, one-way analysis of variance, Tukey’s post hoc test. *P<0.05, **P<0.01, ***P<0.005. All error bars indicate s.e.m.
