Fig. 1

Piezo1 channel leads to iron overload under mechanical stress. a Schematic illustration of mechanical stimulation devices. b Schematic illustration of the cells treated with different drugs under 1 MPa mechanical stress (n = 3). c Heatmap illustrating the different genes expression in NPCs. d Volcano plot of differentially expressed genes in 1 MPa mechanical stress. e, f GO a KEGG bubble plots of differentiated pathways in 1 MPa mechanical stress. g GSEA analysis showing the changes in regulation of metal ion transport and lipid pathway. h Detection of intracellular Fe²⁺ in NPCs at different time-points after stress treatment using FerroOrange and quantitative analysis of relative mean fluorescence intensity (MFI) (n = 3). i Western blot analysis of iron metabolic markers (TFRC, FPN, DMT1 and FTH1) and ferroptotic markers (ACSL4, FSP1, and GPX4) in different time after 1 MPa mechanical stimulation (n = 3). j Intracellular Fe²⁺ content are measured by Iron Assay Kit at different time-points (n = 3). k NPCs were treated with GsMTx4 or Fer-1 under 1 MPa mechanical stimulation for 6, 12, 24, 36, 48, and 72 h (n = 3). l The morphology of rat NPCs treated with GsMTx4 or Fer-1 under 1 MPa mechanical stimulation. m Observation of cytoskeleton in NPCs using Actin Tracker Kit. n, o Quantitative analysis of relative intracellular Fe²⁺ and MDA content of NPCs treated with 1 MPa mechanical stress with or without GsMTx4 for 24 h (n = 3). p Detection of intracellular Fe²⁺ in NPCs treated under different treatments for 24 h using FerroOrange and quantitative analysis of relative MFI (n = 3). q, r Cell death/live analysis showing cell death ratio of NPCs (n = 3). s, t Representative morphological changes in AFCs, BMSCs, and MC3T3-E1 after treatmen twith or without GsMTx4 under 1 MPa mechanical stress (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