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. 2009 Apr 1;20(7):2041–2048. doi: 10.1091/mbc.E08-07-0699

Figure 5.

Figure 5.

Knockdown of Survivin by shRNA increases FKHRL1-induced apoptosis in SH-EP/FKHRL1 cells. (A) The pQ-tetH1-shSurvivin-SV40Puro retrovirus vector was infected into SH-EP/FKHRL1 cells. Knockdown of endogenous Survivin was analyzed by immunoblot. (B) SH-EP/FKHRL1-shCtr (infected with the empty plasmid pQ-tetH1-SV40Puro) and SH-EP/FKHRL1-shSurv cells were treated with 20 nM 4OHT for 24 and 48 h. Apoptosis induction was determined by PI-FACS analysis. (C) Mitochondrial activity was assessed by CMX-Ros staining in untreated and 4OHT-treated (20 nM) SH-EP/FKHRL1-shCtr and SH-EP/FKHRL1-shSurv cells after 48 h. Shown is the mean of three independent experiments. (D) SH-EP/FKHRL1 cells were treated either with 20 nM or with 75 nM 4OHT for 0, 8, or 24 h, and the expression of Survivin and p27Kip1 was determined by immunoblot. GAPDH served as loading control. Densitometry was performed using LabWorks 4.6 software.