Stat3 60199 1 ig

Antibodies specific for CD147 (sc-9754), α-tubulin (sc-8035), WAVE2 (sc-10392), donkey anti-goat IgG-FITC (sc-2024) and FAK Inhibitor 14 (Y-15) (sc-203950) were purchased from Santa Cruz (Dallas, Texas); phospho-FAK (Tyr397) (3283), phospho-Src (Tyr416) (2013), phospho-myosin light chain 2 (Thr18/Ser19) (3674), Src (2109), and myosin light chain 2 (3672) antibodies were purchased from Cell Signaling Technology (Boston, MA); DOCK8 (11622-1-AP) and STAT3 (60199-1-Ig) antibodies were obtained from Proteintech (Wuhan, China); phospho-STAT3 (Tyr705) (ab76135) antibody was obtained from Abcam (Cambridge, UK); phospho-Src (Tyr527) (orb14869) antibody was obtained from Biorbyt LLC (San Francisco, CA); and Alexa 594-conjugated goat anti-mouse IgG and Alexa Fluor 488 phalloidin were purchased from Invitrogen (Carlsbad, CA). The Src kinase inhibitor Src I-1, FAK inhibitor PF573,228 (Sigma, St. Louis, MO) and STAT3 inhibitor WP1066 (Merck Millipore, Darmstadt, GER) were also used. Src I-1 was used at 300 nM, PF573,228 at 300 nM, Y-15 at 10 μM and WP1066 at 5 μM. The Cell Proliferation Reagent WST-1 was purchased from Roche (Mannheim, GER).

Cells were directly collected, and whole cell lysates (WCLs) were lysed in NP40 buffer supplemented with 20 mM β-glycerophosphate and 1 mM sodium orthovanadate. The following primary antibodies and reagents were used: Stat3 (60199-1-Ig, Proteintech, Rosemont, IL, USA), p-Stat3 (Tyr705) (YP0251, Immunoway, Plano, TX, USA), CD44 (60224-1-Ig, Proteintech, Rosemont, IL, USA), p-CD44 (Ser706) (YP0349, Immunoway, Plano, TX, USA), E-cadherin (YT1454, Immunoway, Plano, TX, USA), N-cadherin (YT2988, Immunoway, Plano, TX, USA), Snail (YT4351, Immunoway, TX, Plano, TX, USA), Claudin-1 (YT0942, Immunoway, Plano, TX, USA), ACTB (A5441, Sigma-Aldrich, St. Louis, MO, USA), Afatinib (S1011, Selleckchem.com, Houston, TX, USA), IL-6 (90107ES08, Yeasen, Shanghai, China).