Phospho lats1 thr1079

Antibodies against YAP, phospho-YAP Ser127, TAZ, LATS1, phospho-LATS1 Thr1079, ERK1/2, and phospho-ERK1/2 were purchased from Cell Signaling Technology (USA). Anti-β-actin was obtained from Abfrontier (Korea). To detect the active form of RhoA, the Active Rho Detection Kit (#8820S) was utilized (Cell Signaling Technology).
Immunoblotting was performed as follows: cells were harvested with lysis buffer (0.1% SDS, 50 mM Tris-HCl pH7.5, 150 mM NaCl, 1% NP-40, 0.25% sodium deoxycholate) containing 1× protease inhibitor cocktail (P3100-001; GenDEPOT, USA) and 1× phosphatase inhibitor cocktail (P3200-001; GenDEPOT). Lysates were centrifuged at 18,000g for 20 min prior to boiling with Laemmli SDS sample buffer for 10 min, subjected to SDS-PAGE and transferred to PVDF membranes, which were subsequently blocked for 30 min in 2% bovine serum albumin (BSA) in Tris-buffered saline containing 0.1% Tween 20 and probed with the relevant primary antibodies, followed by horseradish peroxidase (HRP)-conjugated secondary antibody for detection of protein bands.