Fig. 7

Evidence of TGFBR1 activation in ovarian granulosa cells of TGFBR1-CA^G9Cre mice. a-d Real-time PCR analysis of expression of TGFBR1 ^CA, Smad7, Inha, and Zp3 in ovaries from control and TGFBR1-CA^G9Cre mice at PD3 and PD7. Real-time PCR was performed using ΔΔCT method. Data are mean ± s.e.m. n = 4-5. ^* P < 0.05, ^** P < 0.01, and ^*** P < 0.001. Ns, not significant. e Western blotting analysis of TGFBR1^CA, phospho-SMAD2/3, and HSD3B using ovaries from 2-month-old control and TGFBR1-CA^G9Cre mice. TGFBR1^CA was detected using an anti-HA antibody. n = 3. Each lane represents an independent sample. f-i X-gal staining using ovaries from Rosa26/Gdf9-iCre mice (f-h) and Rosa26 control mice (i). Panels g and h are higher magnification images of two different fields of panel (f). At least 3 independent samples per group were used. j Real-time PCR analysis of the expression of Gli1, Gli2, and Tgfbr3 using ovaries from 8-week-old control and TGFBR1-CA^G9Cre mice. Data are mean ± s.e.m. n = 4-5. ^* P < 0.05 and ^*** P < 0.001. k-n RNAscope in situ hybridization analysis of Gli1 mRNA distribution using 8-week-old control (k) and TGFBR1-CA^G9Cre ovaries (l). n = 4. Positive and negative controls using TGFBR1 ^{CA flox/+} ovaries were shown in (m) and (n), respectively. Sections were counterstained with hematoxylin. Scale bar is representatively shown in (f) and equals 25 μm (g, h, and k-n) and 100 μm (f and i)