Remifentanil preconditioning protects against hypoxia-induced necroptosis in HCM cells. (A, B) Western blot analysis of the levels of RIP1, phospho-RIP1, RIP3, phospho-RIP3, MLKL and phospho-MLKL. Data were normalized to β-actin. The levels of phospho-RIP1, phospho-RIP3 and phospho-MLKL are also presented as a ratio of phospho-RIP1 to RIP1, phospho-RIP3 to RIP3 and phospho-MLKL to MLKL, respectively. (B) Bars indicate SD, n = 3, ***p < 0.001, **p < 0.01 compared to normoxic control (CTR), ###p < 0.001 compared to hypoxic control (CTR) (ANOVA and Dunnett's a posteriori test). CTR, control; R, remifentanil preconditioning.