Figure 4. PlxnA2 GAP Forward Signaling Is Required for Proper Distribution of GCs in the Developing DG.
(A and B) Western blot analysis of mouse forebrain cell-culture lysates. (A) Blots of total cell lysates of Plxna2+/+, Plxna2R/R, Plxna2+/−, and Plxna2−/− DIV10 (10 days in vitro) cultures and (B) biotinylated cell-surface proteins isolated from Plxna2+/+ and Plxna2R/R cultures probed with anti-PlxnA2, anti-TfR, TuJ1, t-PlxnA2 (total PlxnA2), or s-PlxnA2 (surface PlxnA2).
(C) Quantification of t-PlxnA2 in Plxna2+/+ and Plxna2R/R lysates normalized to TuJ1 (left; n = 6) and s-PlxnA2 normalized to s-TfR (right; n = 4). Data are shown as mean ± SEM, unpaired two-tailed Student’s t test.
(D) Anti-Prox1 staining of coronal brain sections through the dorsal hippocampus of P1 (A) Plxna2+/+ (n = 5) and Plxna2R/R (n = 6) mouse pups. Scale bar is 200 µm.
(E) Quantification of GC distribution in the Plxna2+/+ (n = 5), Plxna2R/+ (n = 5), and Plxna2R/R (n = 6) DG. Results are shown as mean value ± SD. ***p < 0.001; ****p < 0.0001, one-way ANOVA with Tukey post hoc t test. ns, not significant.