Gfp antibody

The hearts were harvested and fixed with 4% paraformaldehyde and embedded in paraffin. The sections were then immunostained with the rabbit anti–green fluorescent protein (GFP) antibody (GeneTex) and rabbit anti–β‐Gal antibody (Invitrogen). For visualization, a diaminobenzidine substrate kit (Vector Laboratories) was used. To evaluate the degree of fibrosis, Masson's trichrome staining was performed and the infarct size was quantified using the midline length measurement, as previously reported.19 In brief, infarct size was evaluated from 3 sections per sample. To measure the infarct size, the sum of midline infarct lengths from all sections was divided by the sum of midline circumferences from all sections. The infarct size value is presented as a percentage. For immunofluorescent staining of macrophages, the collected hearts were processed for frozen sectioning by embedding in the OCT media. The sections were stained with the mouse anti‐cTnT (DSHB) and rat anti‐F4/80 (Abcam). Appropriate secondary antibodies (Invitrogen) were used for visualization under a fluorescence microscope. Nuclei were stained with 4,6‐diamidino‐2‐phenylindole (1 μg/mL; Sigma) for visualization.