Anti troponin 1

Paraffin‐embedded sections were deparaffinised and then boiled in target retrieval solution (DAKO North America, Inc, CA) for 10 minutes. Staining with either mouse monoclonal anti‐IL‐33 (Alexis Biochemicals, Enzo Life Sciences AG, Lausen, Switzerland, dilution 1:200) or with rabbit polyclonal anti‐Troponin I (abcam, Cambridge, UK, dilution 1:100) antibodies was performed as described.27 The IL‐33 fraction of the stained myocardial tissue was quantified using ImageJ software. Mean fluorescence intensity (MFI) was compared between patients with or without statin treatment in order to determine the myocardial IL‐33 content.

Cells and heart samples were fixed with 4% paraformaldehyde for 10 minutes and permeabilized with 0.2% Triton. After 1 hour of blocking by 10% bovine serum albumin (BSA), the cells or tissue samples were incubated with primary antibodies as follows: anti‐troponin I (Abcam, Cambridge, MA,
http://www.abcam.com). Sections or cells were washed with PBS‐0.05% Tween (PBST) followed by incubation of secondary antibody (fluorescein Isothiocyanate [FITC]‐conjugated; Abcam). After 1 hour of incubation at 37°C, cell nuclei were stained with DAPI. Images were obtained by fluorescence microscope (Leica Microsystems, Wetzlar, Germany,
http://www.leica-microsystems.com) at five randomly selected fields for each sample or well and the mean positive/negative numbers calculated.