2.6.4. Western blot

After the transfected cells grew for 48 h in the 6‐well plate, we washed the cells with 1X PBS three times and lysed them with 100ul 1X RIPA per well. Thereafter, the protein concentration was quantified and 20 μg of proteins from each sample were segregated using 10% SDS‐PAGE gel. The samples were then transferred to PVDF membranes and incubated with primary antibodies (WT1: sc‐7385, Santa Cruz, 1:1000 dilution; GAPDH: sc‐47724, Santa Cruz, 1:1000 dilution) and secondary antibodies (m‐IgGκ BP‐HRP: sc‐516102, Santa Cruz, 1:2000 dilution). Finally, blots were detected with Chemiluminescent HRP Substrate reagent (Merck Millipore, Darmstadt, Germany), and the signal was captured by Bio‐Rad ChemiDoc XRS+ (BioRad Laboratories, Inc., Hercules, CA, USA).