Figure 3

XBP1(S) and HSPA5 mRNA expression levels, and CHOP, phospho-IRE1, phospho-PERK, and TGF-β1 protein expression levels in ovaries of control and PCOS mice. Three-week-old female mice were divided into two groups. The control group (n = 5) was s.c. injected daily with sesame oil for 20 days. The PCOS group (n = 5) was s.c. injected daily with DHEA (6 mg/100 g of body weight) for 20 days. The ovaries were collected on day 21. (A,B) Cross-sections of ovaries from control and PCOS mice were hybridized with a DIG-labeled antisense XBP1(S) or HSPA5 probe. (C–F) Cross-sections of ovaries from control and PCOS mice were stained with an anti-CHOP antibody counterstained with hematoxylin, or an anti-phospho-IRE1, anti-phospho-PERK, or anti-TGF-β1 antibody. (G) Controls for background level (a,b) hybridized with sense probe, (c) stained with isotype IgG, and (d) stained with isotype IgG and hematoxylin. (A–F) (a–d) show the representative sections. Lower panels (c,d) show highly magnified views corresponding to (a,b). (e) show the quantitative analysis of (A,B) in situ hybridization and (C–F) immunohistochemical staining. The scale bars in (A–F) (a,b) and (G) (a–d) indicate 100 μm. The scale bars in (A,B) (c,d) and (C–F) (c,d) indicate 20 μm and 50 μm, respectively. *p < 0.05. NC, negative control; ISH, in situ hybridization; IHC, immunohistochemistry.