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

Figure 5

Iron overload induces the opening of mitochondrial permeability transition pore in osteoblastic cells. The opening of mitochondrial permeability transition pore results in the loss of mitochondrial membrane potential. After exposure to FAC (50-200 μM) for 120 h, the flow cytometry was used to detect the opening of mPTP and changes of MMP. (a) Representative dot plot of the flow cytometry results illustrated the increase of mPTP opening. The quantitative mPTP in osteoblasts is shown by RFI. Values are expressed as the from three independent experiments ( vs. saline control, ANOVA test). (b) Representative dot plot of the flow cytometry results illustrated the decrease MMP. The quantitative MMP in osteoblasts is shown by red/green ratio. Values are expressed as the from three independent experiments ( vs. saline control, ANOVA test). (c) Representative fluorescence images were obtained in situ JC-1 staining by confocal microscopy. . (d) Representative western blots of the total protein expression of PGAM5, DRP1, and mitochondrial protein expression of phosphorylated DRP1 (Ser637). (e) Histogram analysis showing the relative protein levels of PGAM5, DRP1, and mitochondrial protein expression of phosphorylated DRP1 (Ser637). Data are presented as the from three independent experiments ( vs. saline control, ANOVA test).

(a)

(b)

(c)

(d)

(e)