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. 2016 Mar 4;7(15):20080–20092. doi: 10.18632/oncotarget.7914

Figure 2. ABC294640 induces apoptosis and inhibits STAT3 phosphorylation in cholangiocarcinoma cells.

Figure 2

(A) Percentage apoptosis of cholangiocarcinoma cells treated with the indicated concentrations of ABC294640 (20–100 μM) for 72 h. Apoptosis was quantified by assessing characteristic nuclear changes of apoptosis by fluorescence microscopy after DAPI staining. The results are presented as mean ± SEM from at least three independent experiments. (B) WITT cells were treated with 20-60 μM ABC294640 for 72 h. Apoptosis was then measured by Annexin V-FITC/PI labeling followed by flow cytometry. (C) Western immunoblots for PARP and caspases 9, 8 and 3 performed on cell lysates from the same treatment as the flow cytometry assay. (D) Western immunoblots for PARP and caspases 9, 8 and 3 performed on WITT cell lysates prepared after 72 h treatment with ABC294640 in the presence or absence of the pan-caspase inhibitor, Z-VAD-FMK. (E) Cholangiocarcinoma cells were treated with 20–60 μM ABC294640 for 48 h. STAT3 Y705 phosphorylation was determined by immunoblotting.