Figure 5
Apoptosis assay on GSK-J4 treated cell lines. (A) HCT116, HepG2, and SW13 cells were treated with 30 μM GSK-J4 for 20 h and stained with Annexin-V-Alexa Flour 647/Sytox-blue. Apoptosis was analyzed by flow cytometry in triplicate (N = 3) and representative data is shown. The horizontal and vertical axes indicate the Annexin-V and Sytox staining measures, respectively. The cell healthy/apoptotic states are denoted in red. (B) Quantitative representation of the cell healthy/apoptotic states with stacked bar charts. Results of three independent experiments (R1, R2, and R3) are presented. The vertical axis shows the percentage of cells with total cell numbers at 100%. N.A.: data not available. (C) Ratios of late or early + late apoptotic cells to total cells in each cell line are first calculated, and specific ratios were obtained by subtracting ratio(DMSO) from ratio(GSK-J4) and plotted as percentages. Values from each of the triplicate experiments are presented below and indicated by open square, closed circle and open circle, respectively, and the means are presented in parentheses and indicated by red lines. Standard error bars are included. Note that apoptosis induction is low in HCT116 and high in HepG2. Differences in percentages between HCT116 and HepG2 or SW13 are significant (p < 0.05). Also see Supplementary Fig. S1A,B for apoptosis induction by ATF4.
