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. 2016 Mar 8;7(16):22460–22473. doi: 10.18632/oncotarget.7984

Figure 3. Pantoprazole suppresses TOPK activity in vitro and in JB6 Cl41 cells.

Figure 3

A. Pantoprazole inhibits TOPK activity in vitro. The inhibitory effect of pantoprazole on TOPK was determined by an in vitro kinase assay as described in section ‘Materials and methods’. The expression level of phosphorylated histone H3 (Ser10) was confirmed by western blot analysis using an antibody against phosphorylated histone H3. GST-histone H3 was used as a loading control. Data are representatives of results from triplicate experiments. B. Pantoprazole inhibits TOPK activity JB6 Cl41 cells. Cells were treated in the presence or absence of 100μM pantoprazole for various times, then treated with 20 ng/ml EGF for 30 minutes, histones were extracted from cells, electrophoresed on 15% SDS-PAGE gels, total histone H3 and phosphorylated histone H3 proteins were detected by western blot using specific antibodies. Data are representatives of results from triplicate experiments and the intensity ratio of p-Histone H3 to total Histone H3 was calculated by an image J software. C. Different concentrations of pantoprazole were incubated with JB6 cells for 9 h and 2 μM of HI-TOPK-032 was a positive control.