Ab4816

Rat dorsal striatum tissue was homogenized using BioRuptor in 1× IP buffer (150 mM NaCl, 10 mM Tris-HCl pH = 7.4, 1 mM EDTA, 1 mM EGTA, 1% Triton X-100) containing protease and phosphatase inhibitors. Homogenates were pre-cleared using protein G Dynabeads for 1 h at 4 °C. For immunoprecipitation, 1 mg of pre-cleared protein was incubated overnight at 4 °C with 7.5 µg of total Fyn (4023S, Cell Signaling, diluted 1:1000) or 5 µg of total Src (ab16885, Abcam, diluted 1:1000) antibody. Immunocomplexes were pulled down with protein G Dynabeads for 4 h at 4 °C and eluted into Laemmli buffer at 55 °C for 10 min. 50 µg of immunoprecipitated protein was used for western blotting. To determine kinase activity, antibodies against active p(Y418)-Src (ab4816, Abcam, diluted 1:200) and inactive p(Y529)-Src (ab32078, Abcam, diluted 1:1000) were used. In addition, dorsal striatal protein tyrosine kinase activity was also measured using a commercially available kit following the manufacturer’s instructions (Omnia Kinase Assay, KNZ3051, Thermo Fisher).

The following antibodies were used in this study for western blot, immunohistochemistry, immunofluorescence staining and immunoprecipitation: YAP1 (Santa Cruz Biotechnology, sc-376,830, 1:100 dilution for western blot; 1:50 dilution for immunohistochemistry and 1:50 dilution for Immunofluorescence staining; Abcam, ab52771, 1:20 dilution for IP), p-YAP1 (Abcam, ab76252, 1:10000 dilution for western blot), α-SMA (Abcam, ab5694, 1:200 dilution for western blot; 1:100 dilution for immunohistochemistry and 1:100 dilution for immunofluorescence staining), FAP (Abcam, ab53066, 1:1000 dilution for western blot and 1:100 dilution for immunofluorescence staining), SRC (Signalway Antibody, #40790, 1:1000 dilution for western blot, 1:100 dilution for immunohistochemistry and 1:100 dilution for immunofluorescence staining), p-SRC (Abcam, ab4816, 1:1000 dilution for western blot), TEAD1 (Abcam, ab133533, 1:20 dilution for IP and 1:500 dilution for western blot), GAPDH (Sungene Biotech, KM9002, 1:5000 dilution for western blot).