Anti gapdh lf p a0212

EGF, IL-6, and human phospho-kinase array were purchased from R&D Systems (Minneapolis, MN, USA). Other chemicals were purchased from Sigma-Aldrich (St. Louis, MO, USA). The following antibodies were used: anti-DDIAS (HPA038540) from Atlas Antibodies (Stockholm, Sweden); anti-SHP1 (sc-7289), anti-TC-PTP (sc-373835), anti-HA (sc-805), and anti-Myc (sc-40) from Santa Cruz Biotechnology (Santa Cruz, CA, USA); anti-STAT3 (9139, 4904), anti-HA (3724), anti-pSTAT3 (Y705, 9145), anti-pSTAT3 (S727, 9134), anti-pJAK1 (Y1034/1035, 3331), anti-JAK1 (3344), anti-pJAK2 (Y1007/1008, 3771), and anti-JAK2 (3230) from Cell Signaling Technology (Beverly, MA, USA); anti-PTPRM (MAB4446) from R&D Systems; anti-GAPDH (LF-P-A0212) from AbFrontier (Seoul, Korea); and anti-Flag (F1804) from Sigma-Aldrich. Full-length and deletion mutants of Flag-DDIAS were cloned as previously described^{35 (link)}. 4×M67 pTATA TK-Luc (#8688) and Flag-STAT3 Y705F (#24984) were obtained from Addgene (Cambridge, MA, USA). Myc-DDK-tagged PTPRM (RC226398) was obtained from OriGene (Rockville, MD, USA). Full-length and deletion mutants of STAT3-tagged HA were subcloned from the Flag-STAT3 expression vector (Addgene). STAT3 gene was mutated using the QuickChange Site-Directed Mutagenesis Kit (Enzynomics, Daejeon, Korea). The constructs were verified using sequence analysis.

The cells were lysed with RIPA buffer (Millipore, Billerica, MA, USA) containing protease inhibitor cocktail (Roche), and the lysates were quantified with a protein assay kit (Bio-Rad, Hercules, CA, USA). The cell lysates were separated using SDS-PAGE and then transferred to PVDF membranes. The proteins were identified while using appropriate antibodies. Anti-VGLL1 (10124-2-AP) was purchased from Proteintech (Rosemont, IL, USA). Anti-TEAD4 (ab58310) and anti-MMP9 (ab76003) were purchased from Abcam (Cambrige, MA, USA). Anti-GFP (NB600-308) was purchased from Novus Biologicals (Centennial, CO, USA). Anti-Myc (sc-789) and anti-HA (sc-805) were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Anti-GAPDH (LF-PA0212) was purchased from AbFrontier (Seoul, Korea). Anti-β-tubulin (2128), anti-pSer473-AKT (9271), anti-AKT (9272), anti-p-β-catenin (9561), and anti-β-catenin (9562) were purchased from Cell Signaling (Danvers, MA, USA). Anti-Flag (F1804) was purchased from Sigma-Aldrich (St. Louis, MO, USA). The protein signal was detected while using an enhanced chemiluminescence kit (Millipore, Burlington, MA, USA).