Anti ptk2

Proteins were extracted by using RIPA lysis buffer (Beyotime, Jiangsu, China) and the concentrations were detected by using the protein assay kit (Beyotime, Jiangsu, China). Protein lysates were separated by 10% sodium dodecyl sulfate-polyacrylamide gels and then transferred to nitrocellulose membrane (Beyotime, Jiangsu, China), which was incubated 1 h with 5% non-fat milk and immunoblotted overnight at 4 °C with primary antibodies: anti-PCNA(Proteintech, USA), anti-YAP1(Abclonal, China), anti-p-YAP1(Abclonal, China), anti-MST1(Abclonal, China), anti-p-MST1(Sigma, USA), anti-LATS1/2(Abclonal, China), anti-p-LATS1/2(Abclonal, China), anti-PTK2(Abclonal, China), anti-β-actin(Abclonal, China), and anti-LaminA(Proteintech, USA). The next day, the membranes were incubated in secondary antibodies for 1 h at room temperature. Dilutions of all antibodies used in this study were 1/1000. Signals of protein bands were scanned by Odyssey Infrared scanning system (Li-Cor, Lincoln, NE, USA).

Proteins in cells were extracted using RIPA lysis buffer (Beyotime), and concentrations were determined using a protein assay kit (Beyotime). Protein bands were scanned using the Odyssey Infrared scanning system (Li-Cor, Lincoln, NE, USA). The following antibodies were used: anti-PCNA, anti-PTK2 (Abclonal), anti-SRSF1, anti-MMP2, and anti-MMP9 (Santa Cruz Biotechnology). RIP assays were performed using the BersinBio^TM RNA Immunoprecipitation (RIP) Kit (BersinBio), with anti-SRSF1 (Santa Cruz Biotechnology), anti-FLAG (Abclonal), and appropriate control IgG (BersinBio) antibodies. Subsequently, qRT-PCR and agarose gel electrophoresis assays were performed. IHC was performed using antibodies against PCNA, Ki-67, PTK2, and MMP2 (Abclonal). IF was performed using an antibody against SRSF1 (Santa Cruz Biotechnology). Images were taken using a microscope (Leica Microsystems). All assays were performed according to the manufacturer’s protocol. The catalog numbers for all antibodies used were provided in Table S2.