Anti p lats1 t1079

Antibodies against caspase 9 (sc-56077) and β-actin (sc-8432) were obtained from Santa Cruz Biotechnology, Texas, United States. Others were purchased from Cell Signaling Technology: anti-PARP (#9542), anti-cleaved caspase 3 (#9661), anti-MST1 (#3682), anti-MST2 (#3952), anti-p-MST1(T183)/MST2(T180) (#49332), anti-LATS1 (#3477), anti-p-LATS1(S909) (#9159), anti-p-LATS1(T1079) (#8654), anti-MOB1 (#13730), anti-p-MOB1 (T35) (#8699), anti-SAV1 (#13301), anti-YAP (#14074), anti-p-YAP (S127) (#13008), and anti-p-YAP (S397) (#13619). RNase A and Propidium Iodide/PI were purchased from Coolaber (Beijing, China) and Sigma-Aldrich (United States), respectively.

Cells were washed with ice-cold phosphate-buffered saline (PBS) and lysed in 200 μL of RIPA buffer (50 mM Tris-HCl, 150 mM NaCl, 1% sodium deoxycholate, 0.1% SDS, and 2 mM EDTA) containing protease inhibitors and phosphatase inhibitors. The lysates were centrifuged at 16,000 g (Eppendorf, Hamburg, Germany) for 15 min. Subsequently, the cell lysates were analyzed by SDS-PAGE in 8% or 10% gels and transferred to a polyvinylidene difluoride (PVDF) membrane (Millipore, Billerica, MA, USA). Immunoblotting was performed with anti-pYAP, anti-Lats1, anti-pLats1 (T1079), anti-IκB, anti-NF-κB (p65), and anti-pNF-κB (p65) antibodies from Cell Signaling Technology Inc. (Danvers, MA, USA); and anti-YAP, anti-ICAM1, anti-VCAM1, and anti-β-actin antibodies from Santa Cruz Biotechnology (Dallas, TX, USA).