Figure 2.
The coordination of DSCAM and DCC in binding to TUBB3. (A) DCC shRNA or functional blocking anti-DCC antibody inhibited the induction of the interaction of endogenous DSCAM with TUBB3 by Netrin-1. E15 mouse cortical neurons were transfected with Venus YFP only, Venus-YFP plus control shRNA or Venus-YFP plus DCC shRNA and cultured for 2 d. Neurons were stimulated with Netrin-1 or the control in the presence of or absence of anti-DCC functional blocking antibody. (B) Quantification of relative binding of DSCAM and TUBB3. The y-axis shows relative binding of TUBB3 with DSCAM in arbitrary units. Data are mean ± s.e.m from three independent experiments. *** indicates p< 0.001 (One way Anova and Fisher LSD post-hoc comparisons). NS, not significant. (C) DSCAM knockdown blocked the Netrin-1-induced interaction of endogenous DCC and TUBB3. E15 mouse cortical neurons were transfected with Venus YFP only, Venus-YFP plus control shRNA or Venus-YFP plus DSCAM shRNA and stimulated with Netrin-1 or the control. Expression of DSCAM shRNA had no effects on levels of endogenous DCC and TUBB3. (D) Quantification of relative binding of DCC and TUBB3. The y-axis shows relative binding of TUBB3 with DCC in arbitrary units. Data are mean ± s.e.m from three independent experiments. *** indicates p< 0.001 (One way Anova and Fisher LSD post-hoc comparisons). NS, not significant.
