Fig. 4.

Lentiviral shRNA knockdown of hnRNPG leads to ERΔ7 Loss in hTERT-HM cells. RNA and protein total lysates were isolated after hTERT-HM cells were infected with hnRNPG shRNA lentivirus. Representative western blot (A) and densitometric analysis demonstrating a decrease in hnRNPG (B) and ERΔ7 isoform (D) and no change in ERα66 (F) protein levels in hTERT-HM cells upon lentivirus-mediated knockdown of hnRNPG compared to non-silencing pGIPZ shRNA lentiviral control. Q-PCR analysis also demonstrates a knockdown of hnRNPG (C), ERΔ7 (E) and an increase in ERα66/ERα46 (G) mRNA levels upon lentivirus-mediated knockdown of hnRNPG compared to non-silencing pGIPZ shRNA lentiviral control. GAPDH is the loading control. Gene expression was normalized to Rplp0. *p < 0.05 by two-tailed Student's unpaired t-test, each experiment was performed in triplicate with n = 3 per group. Data shown are mean ± SEM.