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Fig. 1. Mlc3f-nlacZ transgene expression status is maintained ex vivo. (A) Expression pattern of the Mlc3f-nlacZ-2 transgene in a neonatal heart, showing ß-galactosidase activity restricted to the right atrial and left ventricular compartments. LA, left atrium; LV, left ventricle; RA, right atrium; RV, right ventricle. (B) Primary ventricular culture following immunocytochemistry to detect -actinin (red) and Hoechst staining to detect nuclei (blue). Most cells in the cultures are -actinin positive cardiomyocytes; a minority of nonmyogenic cells are also present (arrowheads). (C) Co-immunohistochemistry of primary left ventricular cardiomyocytes showing that ß-galactosidase activity (green) is specific to cardiomyocyte nuclei (arrowhead), identified by -actinin expression (red). (D) Phase contrast image of X-gal-stained primary ventricular cultures showing that few ß-galactosidase-positive cardiomyocytes (arrowhead) are observed in right ventricular free-wall cultures, whereas (E) many ß-galactosidase-positive cardiomyocytes are observed in left ventricular free-wall cultures. (F, G) Low magnification bright-field view of right (F) and left (G) primary ventricular cultures after X-gal staining. Scale bars: 10 µm (B-C), 25 µm (D-G).

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