Fig. 6. Disruption of male-specific markers in Pod1 KO gonads. Whole-mount in situ hybridization for Sox9 (A,B) and Dhh (C,D) was performed with gonads from 12.5 dpc embryos. Sox9 and Dhh were expressed by Sertoli cells in wild-type testes (A,C, top), but were absent in wild-type XX gonads (B,D, top). In Pod1 XY mutants (A,C, bottom), expression of both Sox9 and Dhh was reduced relative to controls, especially in the anterior domain of the gonad (arrowheads). Neither Sox9 nor Dhh was expressed in gonads from XX mutants (B,D, bottom). Scc, a marker of Leydig and adrenocortical cells, was expressed in the wild-type XY gonad (E), and in the adrenal primordial cells (ap) of both XY (E) and XX (F) wild-type embryos. Pod1 mutant XY and XX gonads (E,F) had greatly increased Scc expression, particularly at the posterior end of the gonad. Of note, the anterior ends of the Pod1 KO gonads did not separate from the adrenal primordium (E,F; arrowheads), although a distinct boundary between the gonads and adrenal glands was present in the controls (E,F, arrowheads). (G,H) At 11.5 dpc, Scc expression was restricted to adrenal primordia (ap) of wild-type XX and XY embryos, but was seen throughout the gonads (go) within the urogenital ridges of Pod1 mutant XX and XY gonads. (I,J) The adrenal-specific steroidogenic marker 11ß-hydroxylase was expressed appropriately in the adrenal primordia (ap) of both wild-type and Pod1 KO mice at 12.5 dpc. ap, adrenal primordia; go, gonad.