Fig. 4

NCOA4-mediated ferritinophay participated in the acidic-induced ferroptosis in human NPSCs. A, B Representative western blot images of TFR, DMT1, FPN1, and NCOA4 in the acidic-treated NPSCs. The relative band densities of NCOA4 were quantified. C The target prediction of NCOA4 using STRING database. D, E Representative western blot images of FTH1 in the acidic-treated NPSCs. The relative band densities were quantified. F Co-IP analysis of the binding between NCOA4 and FTH1. G Representative western blot images of FTH1, GPX4, SLC7A11, MMP3, and ADAMTS5 in NPSCs treated with 3-MA and CQ under acidic condition. H Western blot analysis showing the expression levels of FTH1, GPX4, SLC7A11, MMP3, and ADAMTS5 after NPSCs treated with NCOA4-siRNA and Control-siRNA under acidic condition. I The expression of NCOA4 and FTH1 and their colocalization in NPSCs were determined by immunofluorescence. Nuclei were stained with DAPI. Scale bar: 50 μm. J NPSCs were labeled with anti-GPX4 antibodies, and representative fluorescence images are shown. Nuclei were stained with DAPI. Scale bar: 50 μm. K Quantification of MDA contents in NPSCs. L Total iron levels in NPSCs. A P-value less than 0.05 was deemed to be statistically significant. (*P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001; ns, not significant)