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. 2021 Sep 3;100(12):101454. doi: 10.1016/j.psj.2021.101454

Figure 5.

Figure 5

Roles of TEAD1 and TEAD3 in the LATS2-induced inhibitory regulation of granulosa cell (GC) proliferation and expression of FSHR, STAR, CYP11A1, ESR1, and ESR2 transcripts. (A) GCs were transfected with reconstructed pYr-adshuttle-4-TEAD1, pYr-adshuttle-4-TEAD3, and pYr-adshuttle-4-TEAD1 and pYr-adshuttle-4-TEAD3 vectors (OE group), pYr-adshuttle-4 empty vector (negative control, NC) and no plasmid (blank control, BC). GCs were transfected with pYr-adshuttle-4-TEAD1, pYr-adshuttle-4-TEAD3, and pYr-adshuttle-4-TEAD1 and pYr-adshuttle-4-TEAD3 vectors for 24 h, and cell proliferation was detected by EdU incorporation assay. (B) Effects of TEAD1 and TEAD3 overexpression on FSHR, STAR, CYP11A1, ESR1, and ESR2 mRNA abundances in the GCs from prehierarchichal follicles (6–8 mm in diameter) was analyzed by RT-qPCR. (C) GCs were administrated with TEAD1/3-specific siRNA (SR group), scrambled siRNA (negative control, NC), and no siRNA (blank control, BC). The cell proliferation transfected with TEAD1 siRNA, TEAD3 siRNA, and TEAD1 and TEAD3 siRNA for 24 h were detected by EdU incorporation assay. (D) Abundances of FSHR, STAR, CYP11A1, ESR1, and ESR2 mRNA expression impacted by TEAD1 and TEAD3 knockdown in the GCs. (E–J) Results of TEAD1 and TEAD3 knockdown by using specific siRNA in the GCs with the LATS2 overexpression. NC, the granulosa cells were transfected with pYr-adshuttle-4 empty vector; OE, the cells were transfected with reconstructed pYr-adshuttle-4-LATS2. Values were represented as mean ± SEM, and bars with superscript symbols indicate a significant difference compared with control groups at ** P < 0.01, * P < 0.05.