Anti actin mab clone c4

To detect protein expression in the esophageal cells, total protein was extracted from the cells by RIPA buffer (150 mM NaCl, 50 mM Tris–HCl [pH 7.5], 1 % Igepal-CA630, 0.5 % sodium deoxycholate, 0.1 % SDS, 50 mM NaF, 1 mM Na3VaO4, and complete protease inhibitor cocktail). Protein expression was analyzed by SDS-PAGE and western blotting with specific antibodies as described previously [40 (link)–41 (link)]. The primary antibodies for protein detection were anti-AXL rabbit polyclonal antibody (pAb) (ab72069, Abcam), anti-phospho-AXL (Tyr702, D12B2) rabbit monoclonal antibody (mAb) (#5724, Cell Signaling Technology), anti-HER2 rabbit antibody (#2242, Cell signaling), anti-phospho -HER2/ErbB2 (Tyr1221/1222, 6B12) pAb (#2243, Cell Signaling Technology), anti-Actin mAb (clone C4, Millipore), anti-ERK 1 (K-23) (Santa Cruz), anti-phospho-p44/42 Erk1/2 (Thr202/Tyr204) mAb (#4370, Cell Signaling Technology), anti-Akt antibody (#9272, Cell Signaling Technology), anti-phospho-Akt (Ser473) (#9271, Cell Signaling Technology), and anti-α-Tubulin (DM1A, Abcam).

The protein expression profiles of cells in response to drug treatment were analyzed by western blotting as described previously (37 (link)). Briefly, total protein was extracted from the cells or tumor tissues by using RIPA buffer and separated by SDS-PAGE followed by western blotting. The primary antibodies for western blotting were anti-AXL polyclonal antibody (pAb) (#8661, Cell Signaling Technology [CST]), anti-phospho-AXL (Tyr702, D12B2, CST) monoclonal antibody (mAb) (#5724, CST), anti-MET mAb (#8198, CST), anti-phospho-MET mAb (#3126, CST), anti-MMP1 mAb (#MAB901, R&D systems), anti-ERK 1 (K-23) pAb (Santa Cruz), anti-phospho-p44/42 Erk1/2 (Thr202/Tyr204) mAb (#4370, CST), anti-Akt pAb (#9272, CST), anti-phospho-Akt pAb (Ser473) (#9271, CST), anti-actin mAb (clone C4, Millipore), and anti-α-tubulin mAb (DM1A, Abcam). Anti-E-cadherin mAb (#14472, CST), anti-vimentin mAb (#5741, CST), and anti-N-cadherin mAb (#13116, CST) were used for analysis of epithelial-to-mesenchymal transition (EMT). The signal intensities were analyzed by ImageQuant 5.1 (Molecular Dynamics, Inc.).