Research Paper Volume 15, Issue 13 pp 6011—6030

Figure 5. Inhibition of BUB1 kinase activity by 2OH-BNPP1 prolongs EGFR half-life. (A) A549 lung adenocarcinoma cells were treated with 2OH-BNPP1 (10 μM) and Cycloheximide (50 μg/ML) for 1 hour followed by EGF (50 ng/mL). Cells were harvested at different time points after EGF treatment and resulting lysates were run on SDS-PAGE gels and probed with EGFR, Her2 and Actin antibodies. (B) Densitometric analysis of EGFR blots in A549 was performed using ImageJ. Resulting data was analyzed in MS-Excel. The protein half-life plots (t1/2) were generated using GraphPad Prism. The plots are of combined data from 2-3 biological repeats is shown. (C, D) NCI-H358 cell lines was treated similar to A549 and EGFR protein half-life was estimated by densitometric analysis.