Fig. 7

Vegf signaling is required for the survival of deep-layer neurons and proper cortex formation. Vegf expression level was quantified using RT-qPCR (A). The mRNA levels of Vegf mRNA normalized to that of Gapdh. The relative values are presented as a bar graph. White bar, WT; gray bar, KO. Data are the mean ± S.E.M of 3 embryos. Statistical differences were assessed using Student’s t-test, * p < 0.05. pCI (control) or sFlt1-pCI (sFlt1) plasmid was electroporated into neural progenitor cells at E13.5 (B–E, G–N). Apoptotic cells (magenta) and transfected cells (green) are detected with anti-cleaved caspase3 and anti-EGFP antibodies, respectively, in coronal sections of the cortex at E18.5 (B–E). The total number of cleaved caspase3+ cells in the cortex (500 μm-wide region) is presented as a bar graph (F). White bar, control; gray bar, sFlt1. Data are the mean ± S.E.M of five sections. Statistical differences were assessed using Student’s t-test, * p < 0.05. Indicated cortical neurons (magenta) and transfected cells (green) are detected with anti-Satb2 (G, H, K, L) or anti-Ctip2 (I, J, M, N) and anti-EGFP antibodies, respectively, in coronal sections of the cortex at E18.5. Abnormal localization of cortical neurons is indicated with the asterisk. sFlt1-pCI plasmid was electroporated into neural progenitor cells at E15.5 (O–T). Apoptotic cells (magenta) and transfected cells (green) with anti-cleaved caspase3 and anti-EGFP antibodies, respectively, in coronal sections of the cortex at E18.5 (O and P). Indicated cortical neurons (magenta) and transfected cells (green) with anti-Satb2 (Q and R) or anti-Ctip2 (S and T) and anti-EGFP antibodies, respectively, in coronal sections of the cortex at E18.5. Scale bars, 100 μm (B–E, G–T). Three (B–E, O–T) and four (F–M) independent experiments are performed and one representative image is shown, respectively