Cells were lysed using Radioimmunoprecipitation Assay Buffer (RIPA) supplemented with a protease inhibitor cocktail (Sigma-Aldrich). Protein concentrations were determined using the Bio-Rad Protein Assay Kit (Bio-Rad, Hercules, CA, USA). Protein samples were separated by 12% sodium dodecyl sulphate‐polyacrylamide gel electrophoresis and transferred onto nitrocellulose membranes. The membranes were incubated with primary antibodies recognizing CD133 (ab19898, Abcam; 1:1000 dilution), SIRT6 (1:500 dilution) or GAPDH (ab181602, Abcam; 1:5000 dilution) overnight at 4 °C, followed by a horseradish peroxidase-conjugated secondary antibody (Sigma-Aldrich). The protein bands were visualized using the ECL Plus Chemiluminescence Detection Kit (Thermo Fisher Scientific, Rockford, IL, USA).

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