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. 2021 Oct 27;2021:3027954. doi: 10.1155/2021/3027954

Figure 3.

Figure 3

EtOH induces osteoblast necroptosis by activating the RIPK1/RIPK3/MLKL signaling in vitro. Western blot showed that the expressions of RIPK1 and p-RIPK1 (a, b), RIPK3 and p-RIPK3 (c, d), and p-MLKL (e, f) on MC3T3-E1 cells were elevated in the EtOH treatment group, while the expression of caspase-8 was not significantly influenced (g, h). Double-labeled immunofluorescence staining confirmed that necroptosis makers RIPK1 and RIPK3 coexpressed in the EtOH-treated MC3T3-E1 cells (i). Bar: 15 μm. And p-MLKL aggregated on the cell membrane, leading to perforation and destruction of the integrity of proliferating MC3T3-E1 osteoblasts (j). Bar: 20 μm. Error bars represent the SD from the mean values. ∗p < 0.05. p-MLKL: phosphorylated mixed lineage kinase domain-like protein.