Fig. 3.

Pharmacological activation of the Piezo1 channel significantly increases iron overload. a Schematic illustrations of cells treated with Piezo1 agonist Yoda1 or inhibitor GsMTx4 under high iron environment. RNA sequencing analysis in rat NPCs treated with 10 μmol/L Yoda1 for 24 h in a Ca²⁺-free medium (n = 3). b Circle map analysis of NPCs. c Volcano plot of differentially expressed genes of NPCs treated with yoda1. d Microarray heatmap illustrating the different genes expression in NPCs. e, f GO and KEGG bubble plots analysis in NPCs. g GSEA analysis showing the changes in regulation of response to mechanical stimulus and iron ion homeostasis pathway. h Chemical structure of FAC. i Representative morphological changes of NPCs treated with Yoda1 or GsMTx4 in 100 μmol/L FAC for 24 h. j Actin Tracker Kit showing the changes of cytoskeleton in NP cells. k Detection of intracellular Fe²⁺ using FerroOrange and quantitative analysis of relative MFI (n = 3). l, m Quantitative analysis of relative intracellular Fe²⁺ and MDA content of NP cells treated with or without Yoda1 or GsMTx4 for 24 h in high iron environment (n = 3). n The cell death ratio was tested and quantification by cell death/live analysis (n = 3). o NPCs were treated with different drugs for 6, 12, 24, 36, 48, and 72 h, cell viability was assayed by CCK8 Kit (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