Rabbit anti lats2

At the end of the incubation periods, islets and INS-1E cells were washed in ice-cold PBS and lysed in RIPA lysis buffer containing 50 mM Tris HCl, pH 8, 150 mM NaCl, 1% NP-40, 0.5% sodium deoxycholate, and 0.1% SDS supplemented with Protease- and phosphatase inhibitors (Pierce, Rockford, IL, USA). Protein concentrations were determined with the BCA protein assay (Pierce). Equivalent amounts of protein from each treatment group were run on a NuPAGE 4–12% Bis-Tris gel (Invitrogen) and electrically transferred onto PVDF membranes. After blocking by 2.5% milk (Cell Signaling) and 2.5% BSA, membranes were incubated overnight at 4 °C with rabbit anti-cleaved caspase-3 (#9664), rabbit anti-PARP (#9532), rabbit anti-cleaved PARP (rat specific #9545), rabbit anti-phospho YAP(S127) (#4911), rabbit anti-LATS2 (#5888), rabbit anti-tubulin (#2146), rabbit anti-GAPDH (#2118), rabbit anti-β-actin (#4967) (all Cell Signaling Technology), and rabbit anti-PDX1 (#47267) and rabbit anti-p-MST1 (#79199) (both from Abcam) antibodies, all at a dilution of 1:1000, followed by horseradish-peroxidase-linked anti-rabbit IgG (Jackson). Membrane was developed by using a chemiluminescence assay system (Pierce) and analyzed using DocIT^®LS image acquisition 6.6a (UVP BioImaging Systems, Upland, CA, USA). Uncropped and unprocessed scans of all Western blots are available in the Source Data file.

Cells were harvested in EBC lysis buffer (50 mM Tris-HCl at pH 7.5, 150 mM NaCl, 5 mM EDTA, 0.5% NP-40), supplemented with Complete Mini Protease and Phosphatase Inhibitor Cocktails (Roche, Indianapolis, IN, USA). Cells were lysed and 30-80 μg protein subjected to SDS-PAGE followed by transfer onto an Immobilon-FL PVDF membrane (Millipore, Billerica, MA, USA) and incubation with the indicated antibodies. Detection was carried out with an Odyssey Infrared Imaging System (LI-COR Biosciences, Lincoln, NE, USA) with IR dye-tagged secondary antibodies (LI-COR Biosciences). The following antibodies were utilized: mouse anti-YAP, goat anti-NF2, mouse anti-AMOT, goat anti-AMOTL1, goat anti-AMOTL2 (Santa Cruz, Dallas, TX, USA), mouse anti-FlagM2 (Sigma, Saint Louis, MO, USA), rabbit anti-LATS1, rabbit anti-LATS2, rabbit anti-p-YAP (Cell Signaling, Danvers, MA, USA), TNKS1/2 (Santa Cruz, Dallas, TX, USA), mouse anti-TEAD4, mouse anti-RAS (Thermo Scientific, Waltham, MA, USA), mouse anti-α-Tubulin, mouse anti-β-actin (Sigma, Saint Louis, MO, USA).