ROS-Mediated Necroptosis Is Involved in Iron Overload-Induced Osteoblastic Cell Death

Figure 7

ROS are required for iron overload-induced necroptosis in osteoblastic cells. Osteoblasts were treated with 200 μM FAC with or without NAC (1 mM) for 120 h. (a) Representative graphs were obtained from a flow cytometer after H2DCF-DA staining in osteoblasts. Histogram statistical analysis illustrated the production of ROS in osteoblasts. Data are expressed as the from three independent experiments ( vs. saline control, ^# vs. FAC 200, ANOVA test). (b) Representative data were analyzed using a flow cytometer after Annexin-V/PI double staining in osteoblasts. Histogram analysis demonstrating the ratio of PI positive cells. Values are expressed as the from three independent experiments ( vs. saline control, ^# vs. FAC 200, ANOVA test). (c) Representative western blots of the expression of phosphorylated RIPK1, RIPK3, MLKL, and corresponding total protein. (d) Histogram analysis showing the relative protein levels of phosphorylated RIPK1, RIPK3, MLKL, and corresponding total protein. Data are presented as the from three independent experiments ( vs. saline control, ^# vs. FAC 200, ANOVA test).

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