Ab70562

Antibodies used for WB and immunofluorescence: anti-YAP is sc101199 (1:1,000, Santa Cruz Biotechnology), anti- GR is BK3660S (1:1,000, Cell Signaling), anti-actin is C11 (1:2,000, Sigma), anti-ANKRD1 is 11427-1-AP (1:1,000, Proteintech (DBA), anti-pYAP (Ser127) is 4911S (1:1,000, Cell Signaling), anti-WWTR1 is HPA007415 (1:1000, Sigma), phalloidin is A12379 (1:250, Alexa Fluor), anti-PARP-85 is TB273 (1:500, Promega), anti-vinculin is V4505 (1:4,000, Sigma), anti-LATS1 is ab70562 (1:500, Abcam), phospho-LATS1 (Thr1079) is BK8654S (1:500, Cell Signaling), anti-FAK (C-20) is sc-558 (1:1,000, Santa Cruz), anti-FAK (phospho Y397) is ab81298 (1:1,000, Abcam), anti-Src is BK2110S and anti-Phospho-Src (Tyr416) is BK2101S from Cell Signaling (1:1,000), anti-FN1 is GTX112794 (1:1,000, GeneTex), anti-Slug is C19G7 (1:500, Cell Signaling), anti-Mst1 is 3682S (1:500, Cell Signaling) and anti-phospho-Mst1/2 (T183/T180) is 3681S (1:500, Cell Signaling).

Total protein was prepared using the RIPA buffer (Invitrogen, Thermo Fisher Scientific, Waltham, MA, USA). The proteins were separated by 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis and transferred to polyvinylidene fluoride membranes. Skimmed milk (5%) was blocked at room temperature for 1 h. Next, the membranes and primary antibodies were incubated overnight at 4°C. They were then incubated with secondary antibodies at room temperature for 1 h. Finally, the proteins were identified using enhanced chemiluminescence reagents (Thermo Fisher Scientific) using an enhanced chemiluminescence detection system. GAPDH served as a loading control for total proteins. The primary antibodies used were as follows: TRIM65 (ab165884, Abcam, Cambridge, MA, USA; 1 : 1000 dilution), LATS1 (ab70562, Abcam, Cambridge, MA, USA; 1 : 1000 dilution), LATS2 (ab110780, Abcam, Cambridge, MA, USA; 1 : 1000 dilution), GAPDH (ab181602, Abcam, Cambridge, MA, USA; 1 : 5000 dilution), and ubiquitin (ab7780, Abcam, Cambridge, MA, USA; 1 : 1000 dilution).