Figure 6: Dhh and Ihh are required for theca cell development in the ovary.

(a–d) Immunofluorescence of α-SMA in red (a, b), CYP17A1 in magenta (c, d) and FOXL2 in green in Dhh/Ihh DKO (n=3) and control ovaries (n=7). α-SMA is a marker for smooth muscle cell. CYP17A1 is a marker for androgen-producing theca cells and FOXL2 marks granulosa cells. Scale bar, 200 μm. (e) Quantitative PCR analysis of gene expression for Hedgehog signalling components (Dhh, Ihh and Gli1), and genes associated with steroidogenesis (Nr5a1, Star, Cyp11a1 and Hsd3b1) in control (n=4) and DKO ovaries (n=3). Two-tailed Student’s t-test was used. *P<0.05; **P<0.01. (f) Serum levels of DHEA, testosterone and progesterone in control (n=7) and Dhh/Ihh DKO female mice (n=3). *P<0.05. Two-tailed Student’s t-test was used and values in all graphs are presented as means±s.e.m.