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. 2022 Feb 10;10:810327. doi: 10.3389/fcell.2022.810327

FIGURE 4.

FIGURE 4

Blockade of GCH1/BH4 sensitizes erastin-induced ferroptosis by activating ferritinophagy. (A) Western blot analysis of GCH1, xCT, GPX4, NCOA4, FTH1, LC3B, and GAPDH in HCT116 and HT29 cells treated with 40 µM erastin for 24 h. The numbers below the LC3 lane indicate the ratio of LC3B-II/LC3B-I. (B) Autophagic flux was determined by the accumulation of LC3B-II in a 4-h treatment period with 25 nM bafilomycin A1 (BafA1). (C) Cell viability after pretreatment with 3-methyladenine (3-MA) for 24 h and treatment with erastin for 24 h in HCT116 and HT29 cells. (D) Western blot analysis of GCH1, FTH1, GPX4, and GAPDH in HCT116 and HT29 cells treated with RSL3 for 24 h. The error bars represent standard deviation from at least three replicates (**p < 0.01, ****p < 0.0001, ns, not significant, compared between the two groups by unpaired t-test).