Figure 3

Effects of STK33-RNAi and PD98059 on Fadu cells and in vitro cell function test. (A) Cells with GFP signal were considered successfully transfected. Increase of the green fluorescence in Fadu cells transfected with both STK33-RNAi and scrambled RNAi was correlated with the post-transfection time, however, apoptosis (↑) was induced only in the cells subjected to STK33-RNAi (Magnification × 400). a) The scrambled RNAi, 1 × 10⁷ TU/ml, b) The STK33-RNAi, 1 × 10⁷ TU/ml. (B) Transfection of STK33-RNAi into the cells led to a significant decrease in STK33 protein expression compared with that in the mock-transfected cells. a) The scrambled RNAi, 1 × 10⁷ TU/ml, b) STK33-RNAi, 1 × 10⁷ TU/ml, c) STK33-RNAi, 1 × 10⁸ TU/ml. (C) Data represent the mean ± SEM from three western blot analyses * P < 0.05.(D, E) MTT assay showed that the decrease in cell viability with PD98059 appeared in a concentration- and time-manner and the decrease in cell viability with STK33-RNAi was correlated with the post-transfection time. The effect exerted by STK33-RNAi was enhanced by PD98059. In addition, induction of apoptosis (↑) and reduction of colony size in Fadu cells by STK33-RNAi and PD98059 were assessed by Ho.33342 and PI staining (F, Magnification × 200), and colony formation (H, Magnification × 100, Scale bars, 50 μm), respectively. a) Control, b) 5 μM PD98059, c) STK33-RNAi, d) STK33-RNAi and 5 μM PD98059. Quantitative analyses of the number of apoptotic cells (G) and colonies (I) in response to the different stimuli were shown. Data represented the mean ± SEM; n = 3, * P < 0.05.