Anti phospho yap s127

The HEK293, MCF7 breast cancer cells, and A549 lung cancer cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS). HCT116 colon cancer cells were cultured in McCoy’s 5A medium with 10% FBS. Cell detachment was achieved by culturing cells in poly–HEMA (2-hydroxyethyl methacrylate)–coated dishes. Where indicated, the following drugs were used: phosphatase inhibitors (10 mM sodium fluoride, 10 mM sodium pyrophosphate) and 2-DG (25 mM) were from Sigma. Glucose-free DMEM was from Thermo Fisher Scientific. The following antibodies were obtained commercially: anti-CTCF (Cell Signaling Technology, #3418), anti–phospho-RxxS/T motif (Cell Signaling Technology, #9614), anti-LATS1 (Cell Signaling Technology, #3477), anti-MST2 (Cell Signaling Technology, #3952), anti-YAP (Cell Signaling Technology, #14074), anti–phospho-YAP (S127) (Cell Signaling Technology, #13008), anti-Flag (Sigma, #F1804), and anti-tubulin (Sigma, #T9026). Rabbit polyclonal anti–phospho-CTCF (S402) antibodies were custom generated using a specific phosphopeptide (GenScript).