Gsk3β

Western blot analysis was performed as described previously,³¹ The following are used as the primary antibodies: METTL3 (1:1000; Abcam, Cambridge, UK), N‐cadherin (1:10 000; Abcam, Cambridge, UK), αSMA (1:500; Sigma‐Aldrich Corp), ZO‐1 (1:500; Invitrogen, Carlsbad, CA, US), MMP9 (1:500; Abcam, Cambridge, UK), pGSK3β (phospho Ser9) (1:1000, Immunoway Biotechnology, Plano, US), GSK3β (1:1000, Immunoway Biotechnology, Plano, US), cyclinD1 (1:1000, Immunoway Biotechnology, Plano, US), β‐catenin (1:1000, Cell Signaling Technology), β‐actin (1:1000, Cell Signaling Technology) ，β‐tubulin (1:1000, Cell Signaling Technology) and GAPDH (1:1000; Cell Signaling Technology). The following were used as secondary antibodies: goat anti‐rabbit IgG H&L (HRP) pre‐adsorbed antibody (1:5000; Abcam, Cambridge, UK) and goat anti‐mouse IgG H&L (HRP) pre‐adsorbed antibody (1:5000; Abcam, Cambridge, UK).

Proteins were extracted from cells with RIPA lysis buffer (Auragene Bioscience), which was supplemented with a protease inhibitor cocktail (Auragene Bioscience) and PMSF (Auragene Bioscience). Equal amounts (10 μg) of proteins were loaded on SDS‐PAGE and then were transferred to a PVDF Immobilon‐P membrane (Millipore, Billerica, MA). The membrane was blocked with 3% BSA‐TBST at room temperature for 90 min and continuously was probed with indicated primary antibodies at 4°C overnight. Then, the membranes were washed and incubated with specific secondary antibodies 1 h. A β‐actin antibody was used as a control, and the MRP1 (1:500; Abcam), P‐gp (1:400; Proteintech), MGMT (1:800; Abcam), cleaved‐PARP (1:800; Abcam), and GSK3β (1:1000; ImmunoWay) antibodies were used for each group.

Whole‐cell lysates were prepared and Western blot was carried out as recently described.15 The following antibodies were used: PRMT5, β‐tubulin, collagen I (Abcam), E‐cadherin, Vimentin, β‐catenin, p‐EGFR (Y1068), EGFR, (Cell Signaling Technology), Akt, p‐Akt, GSK‐3β, p‐GSK‐3β, p‐EGFR (Y1172), HA (ImmunoWay) and HRP‐conjugated secondary antibodies (Pierce).