Fig. 4.

Conditional deletion of Pdgfra in mouse embryos. Tamoxifen was given to pregnant mice at E9.5 and E10.5, E14.5 embryos were collected to isolate total RNAs of the urorectum for RT-PCR analysis (a) and qPCR analysis (b). Expression of beta-actin RNA was used as internal control. “+” means with RT; “−” means without RT (negative control). Relative expression levels of Pdgfra (mean ± S.E.M.) in control and mutant urorectum were shown and the relative expression level of Pdgfra in controls were taken arbitrarily as one. Western blot analysis was performed on E14.5 urorectum to detect the expression of Pdgfra and beta-actin (b-actin) was used as loading controls (c). E12.5 embryos were collected for immuno-histochemistry (d). Pdgfra expression (brown) was detected in the urorectal mesenchyme of E12.5 control embryos. In contrast, no Pdgfra expression was detected in the urorectal mesenchyme of E12.5 Pdgfra-cKO. gt genital tubercle, ugs urogenital sinus, urs urorectal septum, hg hindgut, cm cloaca membrane. n = number of embryos analyzed