Fig. 4: CISD3 plays a crucial role in the mitochondrial homeostasis.
From: CISD3 inhibition drives cystine-deprivation induced ferroptosis
A Immunofluorescence assays verifying the localization of CISD3 protein in HT1080 cells. CISD3 protein was co-localized with mitochondrial protein Tom20, but not with lysosome marker, Lamp1. B Subcellular localization of the CISD3 protein was analyzed by western blot using protein extracted from purified mitochondrial, cytoplasm, and nucleus fractions. Antibodies against mitochondrial protein Tom20 and nucleoprotein Histone H3 were used as indicated markers. C Representative images of mitochondrial morphology labeled by MitoTracker Red in HT1080 cells with altered CISD3 expressions in the presence or absence of 7.5 μM erastin. Scale bars: 5 μm. D Mitochondrial morphology alteration was monitored through Mitotracker probe staining according to the degree of fragmentation. E The morphological changes of mitochondria were detected by transmission electron microscopy (TEM) in the absence or presence of 7.5 μM erastin. Lower scale bars: 1 μm. F HT1080 cells with scramble or shCISD3 vector were treated with or without 7.5 μM erastin for 8 h, the indicated protein levels were analyzed by western blot. β-actin served as the loading control. G Mitochondrial ROS production was detected by MitoSOX staining and measured by flow cytometry. The values are presented as means ± SD. *P < 0.05, **P < 0.01 versus control or between indicated groups.
