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. 2013 Aug 20;6(8):960–987. doi: 10.3390/ph6080960

Figure 1.

Figure 1

Infection with a lentivirus gene transfer vector encoding Stat5-directed shRNA reduces Stat5 expression and suppresses the growth and viability of human tumor cells. (a) A431 epidermal and PC-3 prostate carcinoma cell lines were infected with lentiviruses encoding either the empty vector (LeGO-G: eGFP), a scrambled shRNA (LeGO-C: mCherry) or pairs of shRNA specific for the mRNA of the human Stat5a and Stat5b isoforms.6 days after infection knockdown-efficiency was verified by western blot, using antibodies recognizing either total or tyrosine phosphorylated Stat5 protein. The detection of total Stat3 served as a control of shRNA specificity. Eight days after infection cell density and morphology was documented by phase contrast and fluorescence microscopy. (b) Over a period of 20 days after viral transduction changes in proliferation and viability of A431 and PC-3 cells were measured by XTT assay at regular intervals. The infection with empty vector and Stat5-shRNA expressing lentiviruses was done in triplicates and the simultaneous infection with scrambled shRNA expressing lentiviruses was done once. Results are shown as the percentage of viable cells compared to the mock-treated control. (c) The same experiments were performed with the T-47D breast and HCT116 colorectal cancer cell line. (n = 3; Ø ± SD) Significantly reduced XTT-values in comparison to empty vector expressing cells are indicated. * p < 0.05, ** p < 0.01, *** p < 0.001 (2-way-ANOVA with Bonferroni correction).