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. Author manuscript; available in PMC: 2020 May 10.
Published in final edited form as: Cell Rep. 2020 Apr 14;31(2):107496. doi: 10.1016/j.celrep.2020.03.060

Figure 4. PGR Suppresses Ptgs2 mRNA Expression in Granulosa Cells by NF-κB Inhibition.

Figure 4.

(A) Localization of Nkbia-expressing cells in UMAP plot for mural granulosa cells. Nfkbia mRNA expression intensity is marked with a gray-red color scale in dots.

(B) Changes in Nfkbia mRNA in isolated granulosa cells in superovulated WT and Esr2-PgrKO. Granulosa cells were isolated from the ovary at 3 h, 6 h, and 9 h after hCG injection and analyzed by real-time PCR. Error bars, SD (n = 4); ***p < 0.001 (Student’s t test).

(C) Quantification of acetyl-p65 protein in granulosa cells of WT and Esr2-PgrKO at 3 h after hCG treatment. Error bars, SD (n = 3); ***p < 0.001 (Student’s t test).

(D) Granulosa cells were isolated from the preovulatory ovary at 48 h after PMSG injection. Granulosa cells were cultured with or without hCG for 3 h and then treated with P4 (progesterone) alone or with RU486 for 3 h. Nfkbia, Ptgs2, and Rpl19 mRNA levels were measured by real-time PCR using specific primers. Error bars, SD (n = 3); *p < 0.05, **p < 0.01, and ***p < 0.001 (ANOVA and Tukey’s post hoc test).

(E) Known transcription binding sites in the promoter of the Ptgs2 gene, including NF-κB binding sites. Graphs indicate the binding of acetyl-p65 to each NF-κB binding site of the Ptgs2 promoter in lysates from granulosa cells under basal conditions and RU486-treated condition. Error bars, SD (n = 3); *p < 0.05 and **p < 0.01 (Student’s t test).