Zfp36

Protein lysates were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and transferred to a polyvinylidene difluoride (PVDF) membrane. The blots were probed with the indicated primary antibodies (β-catenin, BD Biosciences, 1:1000), (Vimentin, BD Biosciences, 1:1000), (Bmi-1, Abcam, 1:1000), (ALDH1, Abcam, 1:1000), (Oct4, Abcam, 1:1000), (Nanog, Abcam, 1:1000), (ABCG2, Abcam, 1:1000), (ZFP36, SANTA CRUZ, 1:1000), (EGFR, Cell Signaling Technology, 1:1000), (p-EGFR, Cell Signaling Technology, 1:1000), (ERK1/2, ABclonal, 1:1000), (p-ERK1/2, ABclonal, 1:1000), followed by HRP (horseradish peroxidase)-labeled secondary antibody. The hybridization signal was detected using enhanced chemiluminescence (ECL) (Cat.No:KGP1122, KeyGEN BioTECH). GAPDH was used as the loading control.

For histology analysis, tumor tissues were fixed with 4% paraformaldehyde (PFA) in PBS, embedded in paraffin, cut into 5mm thick sections, and then deparaffinized, followed by hematoxylin and eosin staining (H&E staining) according to standard procedures. After deparaffinization and rehydration, the paraffinembedded sections were exposed to high pressure for 2 min for antigenic retrieval. The slides were then incubated overnight at 4°C with the indicated primary antibodies (Nanog, Abcam, 1:50), (Ki67, Abcam, 1:50), (ZFP36, SANTA CRUZ, 1:50), (p-EGFR, Cell Signaling Technology, 1:50), (ERK1/2, ABclonal, 1:50).