Subcellular localization of GFP-AnxA6 fusion protein in cardiomyocytes. A. Representative images showing expressions of AnxA6 and its mutants in NRVM. The GFP expression was examined under Olympus (IX71) microscope after 96 hrs of transfection. The GFP-AnxA6 appears to be localized to sarcomeric z-lines (arrow), GFP-AnxA6 ΔN1 is localized to the nucleus (arrow) and GFP-AnxA6 ΔC1 is expressed as aggregate like structures in cytoplasm (arrow), The experiment was repeated with three different batches of myocytes preparation (n = 3). Scale bar represents 10 μm (63×). B. Sarcomeric z-line organization showing striations in AnxA6 knocked down cardiomyocytes. NRVM were transfected with AnxA6 shRNA or scrambled constructs and z-lines were visualized by immunofluorescence with α- actinin antibody after 72 h. Transfected cardiomyocytes were selected based on the GFP expression. Representative images were randomly chosen from a pool of cells drawn from 3 independent experiments. Scale bar represents 15 μm (40×).