Fig. 8

Effects of anti-diabetic drugs on necroptosis in cardiomyocytes. Adult mouse cardiomyocytes were incubated with high glucose (HG) or mannitol (25 mmol/L) in the presence of empagliflozin (EMP, 1 µmol/L), metformin (MET, 500 µmol/L) or Vehicle (Veh) for 24 h. a Representative western blots for phosphorylated RIPK3 (p-RIPK3) and MLKL (p-MLKL) and their total proteins as well as GAPDH. b and c; Quantification of p-RIPK3 and p-MLKL relative to their total proteins, respectively. d and j LDH was measured in culture medium. (e and k) Cell death was assessed by annexin V staining. f Representative western blots for p-RIPK3, p-MLKKL, phosphorylated AMPK (p-AMPK) and their total proteins as well as GAPDH. g–i Quantification of p-MLKL, p-RIPK3 and p-AMPK relative to their total proteins, respectively. Data are mean ± SD, n = 5 different cultures. *P < 0.05 versus Mannitol + Veh and ^†P < 0.05 versus HG + Veh (ANOVA)