FIGURE 4.

4-Hydroxy-2-non-enal induces cell necroptosis in H9c2 cells. (A,B) Necroptotic cells were determined using flow cytometry stained with Annexin V and propidium iodide (PI), and the cell necroptosis ratio was demonstrated as the percentage of Annexin V⁺/PI⁺ cells (n = 5). (C,D) Representative Western blots and relative expression of RIP1, p-RIP1, RIP3, and p-RIP3 under time gradients of 4-HNE treatment in H9c2 cells (n = 5). (E,F) Representative immunoblots and relative expression of RIP1, p-RIP1, RIP3, p-RIP3, MLKL, p-MLKL, Ca²⁺/CaMKII, and p-CaMKII in H9c2 cells treated with different concentrations of 4-HNE (n = 5). *p < 0.05 vs. control group, **p < 0.01 vs. control group.