Fig. 4.

Piezo1 channel-mediated iron overload leads to mitochondrial dysfunction, oxidative stress, and lipid peroxidation. a Representative TEM images of of NPCs exposed to the high iron environment, treated with Yoda1 or GsMTx4 for 24 h. Arrows indicate shrunken mitochondria. b Intracellular ROS detection by DCFH-DA and quantitative analysis of relative MFI. c Mitochondrial function of NPCs evaluated by Mitotracker. d, e WB analysis of mitochondrial functional markers, iron metabolic markers and ferroptotic markers of NPCs. (n = 3). f Quantitative analysis of relative MFI of Mitotracker. g PCR analysis of iron metabolic genes and ferroptotic genes of NPCs treated with different chemical stimulations for 24 h and quantification. (n = 3). h Mitochondrial membrane potential of NPCs was assessed through JC-1 assay. i, j The relative MFI was analysed by ImageJ software (n = 3). k Representative histogram plot for fluorescence of oxidized BODIPY-C11. l Intracellular lipid ROS of NPCs evaluated by C11 BODIPY 581/591. m Iron Assay Kit show the Intracellular Fe²⁺ content of different types of cells treated with different chemical stimulations for 24 h (n = 3). All data are expressed as the mean ± SEM, n = 3 replicates from one representative of 3 independent experiments. ns (no significance), *P < 0.05, **P < 0.01, ***P < 0.001