Hep3B cells were seeded in 6‐well plates at 2 × 105 cells/well. DP7‐C and GalNAc‐Pin1 siRNA complex was incubated with the Hep3B cells at doses of 1 μg GalNAc‐Pin1 siRNA. Hep3B cells were harvested 48 hours later for analysis. Total RNA was extracted with a kit (ForeGene). qPCR reactions were performed on a Bio‐Rad CFX96 system.
The western blot results better showed the silencing efficiency of the siRNA. The protein samples were probed with anti‐GAPDH (Cell Signaling Technology, CST), anti‐Pin1 (CST), anti‐Beta Tubulin (CST), anti‐Lamin B (CST), and anti‐Exportin 5 (CST) antibodies. The cells were incubated with primary antibodies at 4°C overnight. Then, the membranes were incubated with secondary antibodies (CST) for 60 minutes and developed using the ECL detection system (Vazyme).
The western blot results better showed the silencing efficiency of the siRNA. The protein samples were probed with anti‐GAPDH (Cell Signaling Technology, CST), anti‐Pin1 (CST), anti‐Beta Tubulin (CST), anti‐Lamin B (CST), and anti‐Exportin 5 (CST) antibodies. The cells were incubated with primary antibodies at 4°C overnight. Then, the membranes were incubated with secondary antibodies (CST) for 60 minutes and developed using the ECL detection system (Vazyme).