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. 2014 Mar 25;60(3):216–223. doi: 10.1262/jrd.2013-085

Fig. 4.

Fig. 4.

Effects of E2, PPT, DPN, MPP and PHTPP on class 1Igf1 mRNA expression in cultured ovarian granulosa cells. (A) Total RNA was obtained from the cultured ovarian granulosa cells, reverse-transcribed, and amplified by PCR using primers for ERα and ERβ. Each PCR product was electrophoresed and stained with ethidium bromide. (B) Whole-cell extracts were subjected to Western blotting using rabbit polyclonal anti-ERα and anti-ERβ antibodies. (C) Ovarian granulosa cells were treated with E2 (10–9 M), PPT (10–7 M), MPP (10–6 M), DPN (10–8 M) and PHTPP (10–6 M) for 24 h. Igf1 mRNA levels were determined by real-time RT-PCR. Values are means ± SEM of triplicate wells. The expression of each mRNA was normalized for RpL19 mRNA expression. Independent experiments were performed three times, and similar results were obtained. *P < 0.05 and **P < 0.01 compared with the control; #P < 0.05 compared with DPN (10–8 M) alone.