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. 2016 Sep 17;7(46):75155–75164. doi: 10.18632/oncotarget.12090

Figure 4. The kinase activity is required for ASK1 to regulate pancreatic cancer cell proliferation.

Figure 4

A. Western blot analysis of ASK1 phosphorylated at threonine 845 (pASK1) and total ASK1 in PANC1 cells treated with the indicated doses of the ASK1-specific inhibitor NQDI-1. B, C. PANC1 cells were treated with NQDI-1, and cell proliferation was analyzed by SRB (B) and MTT (C) assays. D. Western blot analysis of pASK1 and total ASK1 in AsPC1 cells treated with the indicated doses of NQDI-1. E, F. AsPC1 cells were treated with NQDI-1, and cell proliferation was analyzed by SRB (E) and MTT (F) assays. G. PANC1 cells were treated with the indicated doses of NQDI-1, and the percentage of apoptotic cells was examined by staining with annexin V-FITC and propidium iodide followed by flow cytometry. H, I. Western blot analysis of ASK1 and β-actin expression (H) and examination of cell proliferation by SRB assay (I) in PANC1 cells transfected with the indicated siRNAs and plasmids. KD, kinase dead. J, K. PANC1 cells overexpressing HA, HA-ASK1, or HA-ASK1-KD were subjected to Western blotting (J) and colony formation assays (K). L. Quantification of colonies derived from PANC1 cells transfected with the indicated plasmids. Error bars represent mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001; ns, not significant.