Fig 2: Overexpression of PlxnA4 partially rescues the neuronal migration and callosal projection defects caused by Med13 knockdown.A The over-expression of PlxnA4 is verified by immunostaining PlxnA4 and Flag in the brains of co-transfected cells with the PlxnA4 construct. Arrowheads indicate co-labeled neurons. Scale bar, 50 μm. B Representative images of P7 mouse brains transfected with the indicated constructs at E14.5. The results show that over-expression of PlxnA4 can partially rescue the migration defect in shMed13-transfected neurons. Scale bar, 100 μm. C Representative images of P7 mouse brains show that over-expression of PlxnA4 can largely rescue the callosal projection defect caused by Med13 silencing. Asterisks point to the S1/S2 border region. Scale bar, 100 μm. D Quantification of cell distribution in cortical layers at P7. Results are obtained from 9 sections (three per mouse) for each experiment. One-way ANOVA with post-hoc Turkey test, *p < 0.05 and **p < 0.01, shMed13 or shMed13+PlxnA4 group vs Control. ##p < 0.01, shMed13 vs shMed13+PlxnA4 group. E Quantification of relative axon fluorescence at P7. Results are obtained from 9 sections (three per mouse) for each experiment. One-way ANOVA with post-hoc Turkey test, *p < 0.05 and **p < 0.01, Control vs shMed13. ##p < 0.01, shMed13 vs shMed13+PlxnA4 group. F The knockdown efficiency of shMED13 (human) is verified by Western blot. Student’s t-test, *p < 0.05. G Luciferase reporter assay indicates that Med13 can regulate the transcription of PlxnA4 (n = 3 for each group). Student’s t-test, *p < 0.05.