Figure 4

Effects of ER stress on TGF-β1 and CTGF mRNA expression levels and TGF-β1 secretion by cultured human granulosa-lutein cells. (A–E) Granulosa-lutein cells were treated with tunicamycin at 2.5 μg/mL for 0, 3, 9, or 24 h. (F–K) Granulosa-lutein cells were pre-incubated with TUDCA at 1 mg/mL for 24 h, followed by treatment with tunicamycin at 2.5 μg/mL for 24 h. (L,M) Granulosa-lutein cells were transfected with siRNA (50 nM) or negative control siRNA (50 nM) for 24 h and then, treated with tunicamycin (2.5 μg/mL) for 24 h. (A,F,M) TGF-β1, (B,G) CTGF, (C,I,L) XBP1(S), (D,J) HSPA5, and (E,K) CHOP mRNA expression levels in granulosa-lutein cells were measured by real-time PCR and normalized to that of GAPDH. The values represent means ± SEM of triplicate or quadruplicate experiments, relative to the mean control value. (H) The bioactive TGF-β1 level in cultured cell supernatants was measured by ELISA. The values represent means ± SEM of sextuplicate experiments. The results are representative of at least three independent experiments using three different samples. The letters denote significant differences between groups. *p < 0.05. Tm, tunicamycin; TUDCA, tauroursodeoxycholic acid.