Ab137427

After transfection for 48 hours, the cell protein lysates were extracted with radioimmunoprecipitation assay (RIPA) buffer (Beyotime, Shanghai, China) according to the provided instructions. Proteins were separated by 8% sodium dodecyl sulfate (SDS-)-polyacrylamide gel electrophoresis and transferred to polyvinylidene fluoride membranes. The membranes were then blocked with 5% nonfat dry milk for 1.5 hours at 4°C and rinsed 3 times with Tris-buffered saline with Tween (TBST) for 10 minutes each time. Subsequently, the samples were incubated overnight with primary antibodies against TAZ (1 : 1000, ab119373, Abcam, Cambridge, MA, USA), ANGPTL4 (1 : 1000, ab196746, Abcam), SOX2 (1 : 1000, ab137385, Abcam), CD44 (1 : 2000, ab157107, Abcam), OCT4 (1 : 1000, ab137427, Abcam), and ALDH1A1 (1 : 5000, ab227964, Abcam) at 4°C. Later, the samples were incubated with diluted corresponding secondary antibody IgG (1 : 2000, ab6721, Abcam). After 3 TBST rinsing, the membranes were visualized using enhanced chemiluminescence solution, exposed to X-ray, developed, and fixed, and then, the gray values of target bands were analyzed with gel image processing system software (UVP, Inc., Upland, CA, USA). GAPDH (1 : 2000, ab245355, Abcam) was used as an internal reference.