Anti tgf β

Cells or heart tissue were lysed using RIPA lysis buffer (Beyotime, China). Protein concentration was quantified using a BCA protein-assay kit (Beyotime, China). Protein (30 μg) was separated with 10% SDS-PAGE and then transferred into PVDF membranes (GE Amersham, America). After blocking with 5% skim milk, the membranes were incubated with primary antibodies (anti-ADAM17 1: 1000; anti-phospho-ADAM17 (pThr735), Abcam, USA. anti-collagen I 1: 500, anti- collagenIII 1: 500; anti-TGF-β 1: 1,000; P38 1: 500; pP38 1: 500; ERK1/2 1: 500 p-ERK1/2 1: 300, Wanleibio, China) at 4 °C overnight. Then, membranes were incubated with secondary antibodies at room temperature for 1 h. Protein bands were visualized using BeyoECL Plus (Beyotime, China) and measured using IPP Image-Pro Plus software.

The human ovarian cancer cell lines A2780 and SKOV3 were obtained from the Shanghai Cell Bank of the Chinese Academy of Sciences (Shanghai, China). Both cell lines were cultured in RPMI-1640 medium (Gibco Life Technologies, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (Invitrogen, Carlsbad, CA, USA) at 37 °C in 5% CO2. RIPA was purchased from Sigma–Aldrich (St Louis, MO, USA). Transfections were performed using Lipofectamine 3000 (Invitrogen, Carlsbad, CA, USA). The following antibodies were used: anti-HDAC9, anti-vimentin (Abcam, Cambridge, MA, USA); anti-TGF-β, anti-SMAD2/3, anti-P-SMAD2(S465 + S467)/P-SMAD3(S423 + S425) (Wanleibio, China); anti-E-cadherin, anti-N-cadherin, anti-snail, anti-FOXO1, anti-β-catenin, anti-beta actin, anti-lamin A/C, anti-alpha-tubulin (Proteintech, Chicago, IL, USA); anti-Acetyl-β-catenin (Lys49), anti-slug (Cell Signaling Technology, Beverly, MA, USA).