Ripa protein lysis buffer

Fresh mouse liver tissues were suspended in RIPA protein lysis buffer (Sigma-Aldrich, USA) with protease inhibitor cocktail and homogenized using Tissue Cell-Destroyer DS1000 (Novastar, China). After lysis for 30 ​min on ice, the supernatant was harvested via centrifugation at 4 ​°C, 12,000 ×g for 10 ​min. The protein concentration of liver tissues was determined using BCA Assay Kit (Biosharp, China). Equivalent protein samples were separated by SDS-PAGE and transferred into PVDF membranes (Millipore, USA). After blocked by 5% nonfat milk at room temperature for 1 ​h, the membranes were incubated with the primary antibody overnight at 4 ​°C followed by incubation with the secondary antibody. At last, all protein bands were shown using Immobilon Western Chemiluminescent HRP substrate (Millipore, USA) by a Luminescent Image Analyzer (Syngene, UK). Details of antibodies were listed in Supplementary Table S2.

MTT was purchased from Beyotime Institute of Biotechnology (Shanghai, China); Dulbecco's modified Eagle's medium (DMEM) was obtained from Gibco (Thermo Fisher Scientific, Inc., Waltham, MA, USA); Fetal bovine serum (FBS) was purchased from Hyclone (GE Healthcare Life Sciences, Logan, UT, USA); interleukin (IL)-3, IL-6 and stem cell factor (SCF) were purchased from R&D Systems, Inc., (Minneapolis, MN, USA); TRIzol reagent and Lipofect 2000 were purchased from Invitrogen (Thermo Fisher Scientific, Inc.); SYBR Green PCR Master Mix was obtained from Takara Bio, Inc., (Otsu, Japan); pLenti6.3/V5-DEST lentivirus vector and Packaging Mix were obtained from Invitrogen (Thermo Fisher Scientific, Inc.); RIPA protein lysis buffer was obtained from Sigma-Aldrich (Merck KGaA, Darmstadt, Germany); Antibodies for western blotting including anti-YAP, protein kinase B (AKT1), B-cell lymphoma 2 (BCL2) and BCL2 like protein 1 were bought from Santa Cruz Biotechnology, Inc., (Dallas, TX, USA); apoptosis analysis kits (7-AAD, Annexin V-FITC) were obtained from Nanjing KeyGEN Biotech Co., Ltd., (Nanjing, China) and the ECL-PLUS/kit was obtained from GE Healthcare (Chicago, IL, USA).