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Figure 1 | Cancer Cell International

Figure 1

From: Bim and VDAC1 are hierarchically essential for mitochondrial ATF2 mediated cell death

Figure 1

Role of ATF2 in PTX-induced apoptosis in various tumor cell lines. (A) Control, 100 nM PTX-treated (12 hours treatment), and ionizing radiation-treated (IR, 5 Gy) cells were permeabilized and stained for ATF2. The nuclear localization of ATF2 is indicated by costaining with DAPI (Vector Laboratories, UK). Scale bar: 20 μm. (B) The inhibition of cell viability with PTX or leptomycin B (LMB) and PTX cotreatment was assessed by the (4,5-dimethylthiazol-2-yl)2,5-diphenyl-tetrazolium bromide assay. Data are means ± s.e.m and represent the means from three different experiments. (C) Diagrams of flow cytometry showing the results from the detection of apoptosis with the cells treated as indicated above. (D) Shown is a diagram of three similar FACS apoptosis assay using Annexin V/PI staining for B16F10 cells stably transfected with scrambled or ATF2 shRNA incubation in the absence or presence of PTX for 12 hours. Statistical analysis of apoptosis in the different treatments was performed by ANOVA and t-tests; *P < 0.01. Data are means ± s.e.m (n = 4). (E) Different fractionations purified from the indicated cells were subjected to western blotting with COXIV and β-Actin as the loading control. Representative figures of multiple experiments are shown. The morphometry of p-ATF2 and ATF2 are indicated at right. The columns provide average values of at least 3 independent experiments performed in triplicate; the bars, SE. *P < 0.01 compared with the corresponding control.

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