Figure 2.

Knockdown of endogenous ERRγ inhibits KCNQ1, KCNE1, KCNE3, and KLK1 expression in human placental cells. A, Freshly isolated human cytotrophoblasts were infected with lentiviral vectors carrying shRNA targeting ERRγ (shERRγ) or control shRNA (shcon). Nontreated cells (NTC) served as another negative control. RNA was isolated 72 hours later, and expression of ERRγ, CYP19I.1, K⁺ channel α-subunits (KCNQ1) and β-subunits (KCNE1, KCNE3, and KCNE5) and KLK1 mRNA was analyzed by qRT-PCR; RPLP0 was used as the internal reference. Data are the mean ± SEM of values from 3 independent experiments, each conducted in triplicate, and are expressed relative to expression levels in nontreated cells. *, Significantly different (P < .05) from values of cells transduced with control shRNA. B, Total proteins extracted from cells infected with lentiviral vectors carrying shRNA targeting ERRγ (shERRγ) or control shRNA (shcon) for 72 hours were analyzed by immunoblotting using antisera to ERRγ, KCNQ1, KCNE1, and KLK1 or GAPDH, as loading control.