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Total proteins of BCa cells were extracted by RIPA Lysis Buffer, separated by SDS- polyacrylamide gel electrophoresis (SDS-PAGE) and then transferred onto a piece of polyvinylidene difluoride transfer (PVDF) membrane. Subsequently, all cell membranes were incubated with the following primary antibodies against ADAMTS12(ab203102, Abcam, UK), β-catenin (ab32572, Abcam, UK), p-GSK-3β (sc-373800, Santa Cruz, CA) and GSK-3β (sc-377213, Santa Cruz, CA) and GAPDH (sc-47724, Santa Cruz, CA) at 4°C overnight After incubating with the secondary antibody for 1 hour at room temperature, all bands were measured using an ECL system kit (MultiSciences, Hangzhou, China).

For coimmunoprecipitation, cell lysates were incubated on a rotator with antibody, at 4 °C overnight. The protein antibody protein A/G-dynabead complexes were prepared by adding 30 µL of protein A/G dynabeads (Life Technologies, America), a normal IgG control was assayed simultaneously. After centrifugation to pelletize the dynabeads, the supernatants were subjected to sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS–PAGE) and IB. The immunocomplexes were analyzed by using liquid chromatography-tandem mass spectrometry (LC/MS-MS) or western blot analysis and these are performed as described previously^{40 (link)}. The following antibodies were used in western blot analysis: P-gp (1:5000, ab170904,Abcam, UK), Shc3 (1:250, sc-365598, Santa Cruz Biotechnology, CA), GAPDH (1:500, sc-47724, Santa Cruz Biotechnology, CA), β-catenin (1:5000, ab32572, Abcam, UK), Axin (1:200, sc-293190, Santa Cruz Biotechnology, CA), GSK-3β (1:200, sc-377213, Santa Cruz Biotechnology, CA), H-3 (1:2000, ab1791, Abcam, UK), OCT4 (1:1000, #2750, Cell Signaling Technology, USA), SOX2 (1:1000, #3579, Cell Signaling Technology, USA).