Fig. 3

Promoting PKM2 tetramerization alleviates oxidative stress and apoptosis during cardiomyocyte hypertrophy. A Rat primary cardiomyocytes were treated with 1 μM AVP for 48 h to induce hypertrophy. Scale bar = 200 μm. B The diameter of cardiomyocytes in A was measured. Results are expressed as mean ± SEM, n = 20 cells per group. C–D The ROS level was analyzed by MitoSOX Red. Scale bar = 20 μm. Quantitative results are expressed as mean ± SEM, n = 20 cells per group. E–G The apoptosis of cardiomyocytes was analyzed by flow cytometry. Quantitative results are expressed as mean ± SEM, n = 3. H–J Lysates from primary cardiomyocytes were analyzed by western blotting with indicated antibodies. Relative protein level was quantified and expressed as mean ± SEM, n = 3. K–L As in previous studies, PKM2 tetramerization was analyzed by DSS crosslinking of primary cardiomyocytes. The ratio of tetramer to monomer was quantified and expressed as mean ± SEM, n = 3. M–N PKM2 tetramerization in the right ventricular tissue of control and RVF rats was analyzed by DSS crosslinking. The ratio of tetramer to monomer was quantified and expressed as mean ± SEM, n = 4 rats per group. O–P Rat primary cardiomyocytes were treated with 1 μM AVP for 48 h and 100 μM TEPP-46 for 24 h as indicated. PKM2 tetramerization was analyzed by DSS crosslinking. Q–R The ROS level was analyzed by MitoSOX Red. Scale bar = 20 μm. Quantitative results are expressed as mean ± SEM, n = 20 cells per group. S–U The apoptosis of cardiomyocytes was analyzed by flow cytometry. Quantitative results are expressed as mean ± SEM, n = 3