Supplementary Figure 5: Vav2, but not Vav3, strongly binds to Ret.
From: Identification of a Vav2-dependent mechanism for GDNF/Ret control of mesolimbic DAT trafficking
a, Y2H assays reveal that Vav2 strongly binds to Ret intracellular domain (ICD). In contrast, Vav3 weakly interacts with Ret ICD. b, Co-IP assays show that Ret-flag is immunoprecipitated with Myc-Vav2. c, [3H]DA uptake assays reveal that overexpression of TrkA in HEK293 cells stably expressing YFP-DAT does not significantly alter DAT activity compared to control. (n = 4). One-way ANOVA with Bonferroni's multiple comparisons test: F(2, 9) = 40.37, P < 0.0001; post hoc: t(9) = 6.610, *P = 0.0002. Error bars represent s.e.m. d, Western blotting reveals that treatment of N2a cells with 100 ng/ml GDNF increases the phosphorylation of Ret at Tyr905 site. e, Western blotting reveals that overexpression of Flag-tagged Ret elevates the levels of phosphor-Ret Tyr905 in HEK293T cells. These experiments were repeated at least three times. b, d, e are cropped blot images from their corresponding full-length blots presented in Supplementary Figure 12.
