Fig. 4

Expression of candidate genes is altered in sex-reversed gonads. Whole mount in situ of E9 UGS from male controls, female controls or fadrozole treated females. The expression of male candidate genes BMPR2 (a-c) and ZNF385b (d-f) changed dramatically in fadrozole treated embryos (c, f), where expression was almost at the same level as untreated males (a, d). NZP also had increased expression in treated females (i) compared to untreated (h), although expression of this gene has generally dropped in control males at this stage (g). LAMA1 expression had also dropped in males at E9 (j), and only low expression was found in females (k). A slight increase in expression was seen in treated females (l). CAPN5 was lost from the gonads (dotted lines) but not the adrenal gland of treated females (o) compared to control females (n) . In the gonads it obtained a similar level of expression to control males (m). GPR56, which is absent in males (p), was also lost from treated females (r) whereas in untreated females it is strongly expressed in the left ovary (q). Similarly, fadrozole treatment abolished FGFR3 staining in the gonads (u), to a similar level as untreated males (s) from normal strong expression in females (t)