Fig. 5.

Piezo1-induced iron influx is independent of TFRC. Rat NPCs treated with or without GsMTx4 under 1 MPa mechanical stimulation for 24 h after Tfrc-siRNA transfection. a The morphology of rat NPCs after transfection. b PCR and WB analysis of NPCs and quantitation (n = 3). c Quantitative analysis of relative intracellular Fe²⁺ and MDA. d–f Representative images of NPCs were taken using FerroOrange, DCFH-DA and Mitotracker after transfection (n = 3). g Western blot analysis of TFRC, FPN, DMT1, FSP1 and GPX4 in NPCs treated with mechanical stimulation for 24 h after transfection with si-NC and si-Tfrc (n = 3). Rat NPCs treated with or without the stimulation of Yoda1 or GsMTx4 under a high iron environment and Tfrc-siRNA transfection. h Representative cell morphological changes are shown at 24 h of NPCs. i Relative intracellular Fe²⁺ and MDA were quantified after transfection (n = 3). j–l Representative images of NPCs were taken using FerroOrange, DCFH-DA and Mitotracker. The relative MFI was analysed by ImageJ software (n = 3). m, n Western blot analysis of NPCs treated with mechanical stimulation for 24 h after transfecting and quantitation (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