The autism-linked UBE3AT485A mutant exacerbates Wnt signaling.
A and B, HEK293T cells were transfected with the BAR reporter, CMV-Renilla, and empty vector or the indicated UBE3A plasmid. Cells were serum-deprived and treated with control (L-cell) CM (A) or Wnt3a CM (B). Mean values are shown for firefly:Renilla ratios (n = 12). Error bars represent S.D. Statistical analysis was performed using one-way analysis of variance with Bonferroni post hoc correction. ***, p < 0.0005. C, HEK293T cells were transfected with increasing amounts of plasmid encoding WT UBE3A or UBE3AT485A. Mean values are shown for firefly:Renilla ratios (n = 3). Error bars represent S.D. Statistical analysis was performed using two-way analysis of variance with Bonferroni post hoc correction. ***, p < 0.0005. D, transfected HEK293T cells were grown for 12–16 h in L-cell CM or Wnt3a CM (acute) or in Wnt3a CM for one passage prior to transfection through lysis (chronic). Values are shown as box and whisker plots for firefly:Renilla ratios (n = 6). Whiskers represent the range of maximum and minimum values obtained in our experiments. Statistical analysis was performed using a two-sample t test (two-tailed). *, p < 0.05; ***, p < 0.0005.