Figure 3.

The Effect of Androgen on ISC Proliferation

For counting of BrdU⁺ cells, 10 crypts were randomly chosen for investigation.

(A) For detection of BrdU⁺ cells per crypt in the ORX model, IHC was performed with primary antibody to BrdU. Scale bar, 100 μm.

(B) For detection of BrdU⁺ cells per crypt in the inhibition of AR model, IHC was performed with primary antibody to BrdU. Scale bar, 100 μm.

(C) For detection of BrdU⁺ cells per crypt in the OVX model, IHC was performed with primary antibody to BrdU. Scale bar, 100 μm.

(D) For analysis of active β-CATENIN and total β-CATENIN expression, western blotting was performed.

(E) For detection of intestinal stem cells in the ORX model, IHC was performed with primary antibody to Olfm4⁺. Scale bar, 50 μm.

(F) For analysis of relative mRNA levels of Bmp4, quantitative real-time PCR was performed in the ORX, OVX, and inhibition of AR models.

(G) The phosphorylation of Smad1/5 was assessed by western blot analysis.

Data, indicated as mean ± SD, correspond to three independent experiments (A, C, n = 6; B, n = 5; D, G, n = 2; E, F, n = 4 mice/group/experiment). ^∗p < 0.05, ^∗∗p < 0.01, and ^∗∗∗p < 0.001.