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. Author manuscript; available in PMC: 2018 Feb 15.

Published in final edited form as: Mol Cell Endocrinol. 2016 Dec 8;442:81–89. doi: 10.1016/j.mce.2016.12.002

Fig 5 — Granulosa cells were isolated from immature female rats injected subcutaneously with estradiol (1.5 mg) for three consecutive days. Cells were cultured in serum-free media for 24 hours and then pretreated with AdmiR-122 (1 x 10⁹ pfu) or AdGFP (1 x 10⁹ pfu) for 24 hours. Media was replaced after 24 hours and cells were treated with FSH (50 ng/ml) and E2 (1nM) for six hours. A. Total RNAs were isolated and reverse transcribed, and the resulting cDNAs were subjected to real-time PCR quantitation using specific primers and probes for LRBP (MVK). The graph represents changes in mRNA levels normalized to 18S rRNA and shown as fold-change vs. control. Error bars represent mean ±SE. *p<0.05 vs. AdGFP; **p<0.05 vs. AdGFP+FSH+E2; n=4. B. Gel mobility shift analysis was performed using S100 fractions isolated from different treatment groups containing equal amounts of total protein after incubating with [³²P]-labeled LBS (1.5 x 10⁵ c.p.m). The autoradiogram shown is representative of three independent experiments. Densitometric scanning of the bands is represented in the bar diagram. Error bars represent mean ±SE. *p<0.05 vs. AdGFP; **p<0.05 vs. AdGFP+FSH+E2; n=4.

Fig 5 — Granulosa cells were isolated from immature female rats injected subcutaneously with estradiol (1.5 mg) for three consecutive days. Cells were cultured in serum-free media for 24 hours and then pretreated with AdmiR-122 (1 x 10⁹ pfu) or AdGFP (1 x 10⁹ pfu) for 24 hours. Media was replaced after 24 hours and cells were treated with FSH (50 ng/ml) and E2 (1nM) for six hours. A. Total RNAs were isolated and reverse transcribed, and the resulting cDNAs were subjected to real-time PCR quantitation using specific primers and probes for LRBP (MVK). The graph represents changes in mRNA levels normalized to 18S rRNA and shown as fold-change vs. control. Error bars represent mean ±SE. *p<0.05 vs. AdGFP; **p<0.05 vs. AdGFP+FSH+E2; n=4. B. Gel mobility shift analysis was performed using S100 fractions isolated from different treatment groups containing equal amounts of total protein after incubating with [³²P]-labeled LBS (1.5 x 10⁵ c.p.m). The autoradiogram shown is representative of three independent experiments. Densitometric scanning of the bands is represented in the bar diagram. Error bars represent mean ±SE. *p<0.05 vs. AdGFP; **p<0.05 vs. AdGFP+FSH+E2; n=4.