Figure 3
The defective migration of Bacurd2 siRNA-treated cells is augmented by co-delivery of human BACURD2. (A) Western blotting of lysates from P19 embryocarcinoma cells transiently transfected with control siRNA or Bacurd2 siRNAs, together with an expression construct encoding human BACURD2 as an epitopte-tagged (FLAG) protein. FLAG-BACURD2 protein expression is refractory to Bacurd2 siRNA-mediated knockdown. (B) In utero electroporation studies with E14 mouse brains electroporated with GFP vector and control siRNA (‘control’), GFP vector and Bacurd2 siRNA, and Bacurd2 siRNA with the indicated concentrations of BACURD2 expression construct are indicated. (C) Quantitation reveals that while Bacurd2 siRNA treatment impairs radial migration, co-delivery of 0.4 μg/μl BACURD2 construct restores their migration to control levels, while co-delivery of 1.0 μg/μl BACURD2 construct only partially restores their migration within the embryonic cortex (N > 1,450 cells counted per condition; F 6,45 = 15; P < 0.0001; two-way ANOVA followed by Bonferroni’s post hoc test). (D) An analysis of their intracortical distribution reveals that the defective migration of Bacurd2 siRNA-treated cells is restored with co-delivery of 0.4 μg/μl of BACURD2 construct (N > 500 cells per condition; F 6,42 = 15; P < 0.0001; two-way ANOVA followed by Bonferroni’s post hoc test). Graph plots mean ± SEM. Scale bar represents 100 μm.