Ferroportin-Dependent Iron Homeostasis Protects against Oxidative Stress-Induced Nucleus Pulposus Cell Ferroptosis and Ameliorates Intervertebral Disc Degeneration In Vivo

Figure 3

FPN protected human NPCs against intercellular iron overload and ferroptosis induced by TBHP treatment. (a–e) Scrambled siRNA (siCON) or FPN siRNA (siFPN) transfection was performed before DMSO or TBHP treatment (50 μM) for 12 h. The quantitative values are expressed as (); . (a) Representative western blotting assay and quantitation of the level of FPN and FTL protein, which was normalized to β-actin. (b) Intracellular total free iron levels in human NPCs were assayed using iron assay kit. (c, d) Lipid ROS levels were assayed using C11-BODIPY 581/591 using flow cytometry. (e) Cell viability was examined by the absorbance of CCK-8. (f–j) Lenti-Vector or Lenti-FPN infection was performed before DMSO or TBHP treatment (50 μM) for 12 h. The quantitative values are expressed as (); . (f) Representative western blotting assay and quantitation of the level of FPN and FTL protein, which was normalized to β-actin. (g) Intracellular total free iron levels in human NPCs were assayed using iron assay kit. (h, i) Lipid ROS levels were assayed using C11-BODIPY 581/591 by flow cytometry. (j) Cell viability was examined by the absorbance of CCK-8.

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