2.6. Western blot assays

Whole‐cell lysates of the Panc‐1 and BxPC‐3 cells were prepared using the RIPA buffer (Beyotime, Guangzhou, China) according to the manufacturer's instructions. The protein concentration was determined using a BCA protein assay kit (Pierce, Rockford, IL, USA). The protein lysates were resolved on a 10% polyacrylamide gel with a 5% stacking gel. The proteins were subsequently blotted on polyvinylidene difluoride membranes. The membranes were blocked for 2 h in TBS containing 0.1% (vol/vol) Tween‐20 and 10% (wt/vol) nonfat dry milk powder and then incubated with the primary antibodies overnight at 4 °C. Following the incubation with the secondary HRP‐coupled antibodies for 2 h at room temperature, the membranes were washed with 0.1% TBS/Tween‐20, and the immunocomplexes were detected using the enhanced chemiluminescence kit and a Molecular Imager ChemiDoc XRS System (Bio‐Rad Laboratories, Hercules, CA, USA). β‐Actin was used as the internal loading control.