Embryoid bodies transfected with adenylate kinase isoform siRNA cocktail inhibited in cardiac development.

<p><b>A</b> – Overlay of transmission (gray) and EYFP (green) images of embryoid bodies transfected with siRNA cocktail and EYFP vector during days 4–6 estimated transfection efficiency at ∼50%. <b>B</b> - Transfected cells (green) were found in regions that eventually formed beating areas visualized through increased staining by plasma membrane potential-sensitive RH 237 (red). <b>C</b> and <b>D</b> – Scrambled siRNA did not affect beating areas (<b>C</b>), but the adenylate kinase siRNA cocktail reduced the number and quality of beating areas (<b>D</b>). Significant presence of siRNA transfected cells in beating areas visualized by co-transfection with EYFP (green). <b>E</b> and <b>F</b> - Beating areas visualized using mitochondrial probe JC-1 (green) and RH 237 (red) in control (<b>E</b>) and adenylate kinase siRNA cocktail transfected (<b>F</b>) embryoid bodies. <b>G</b> – Adenylate kinase siRNA cocktail transfection reduced the percentage of beating embryoid bodies compared to no or scrambled siRNA transfections. Creatine supplementation of media, at time of adenylate kinase knockdown, rescued beating. <b>H</b> and <b>I</b> – Contractions of control beating areas and those affected by adenylate kinase siRNA transfection (lower). All scale bars are 20 µm. Representative images and data of n = 6–8.