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. 2023 Mar 15;18(11):2514–2519. doi: 10.4103/1673-5374.371381

Figure 6.

Figure 6

Rapa reduces MPP+-induced PC12 cell ferroptosis via autophagy.

(A) PC12 cells were treated with 0, 2.5, 5, or 10 μM Rapa for 24 hours, and the expression of LC3II and P62 was determined by western blot assay. *P < 0.05, **P < 0.01, vs. 0 (B) Cell viability was measured after treatment with 1 mM MPP+ and/or 5 μM Rapa and/or 5 mM 3-MA for 24 hours by CCK-8 assay. The effect of 5 mM 3-MA on the concentrations of glutathione (GSH; C), malondialdehyde (MDA; D) and reactive oxygen species (ROS; E) in PC12 cells treated with 5 μM Rapa. (F) The effect of 5 mM 3-MA on the protein levels of P62, LC3, recombinant solute carrier family 7, member 11 (SLC7A11), and glutathione peroxidase 4 (GPX4) in 5 μM Rapa-treated PC12 cells. PC12 cells in logarithmic growth phase were taken for experiments. The data presented are from three different passages of cells. The experiments were performed in triplicate. Data are expressed as the mean ± SEM (n = 6–10 per group). *P < 0.05, **P < 0.01 (one-way analysis of variance followed by Tukey’s post hoc test). LC3: Light chain 3; MPP+: 1-methyl-4-phenylpyridinium; Rapa: rapamycin; RSL3: methyl (1S,3R)-2-(2-chloroacetyl)-1-(4-methoxycarbonylphenyl)-1,3,4,9-tetrahyyridoindole-3-carboxylate.