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. 2016 Oct 10;113(43):E6669–E6678. doi: 10.1073/pnas.1603876113

Fig. 5.

Fig. 5.

Glutathione metabolism is impaired on FLT3 inhibition and further impaired by ATM/G6PD inactivation. Metabolomic analysis was performed as described in Fig. 4 by UPLC-MS/MS. (A) Glutathione metabolites in Molm13 cell lines treated with AC220. Asterisks indicate statistical significance (ANOVA; *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001) for comparisons of all samples to vehicle-treated control cells, and pound signs indicate statistical significance (#P ≤ 0.05, ##P ≤ 0.01, ###P ≤ 0.001) for comparisons of ATM or G6PD knockdown cells to control cells under the same treatment conditions. (B) Molm13 cell lines were treated with AC220 (triangle: 0, 1, 2, and 3 nM) for 24 h and glutathione levels were measured using a luminescence based assay (RLU: relative light units; normalized to cell number). (C) Molm13 cell lines were treated with AC220 as in B and glutathione (GSH) levels were measured. (D) Molm13 cell lines were treated with AC220 as in B for 20 h and ROS levels were measured using the fluorogenic dye DCFH-DA (MFI: mean fluorescence intensity) by flow cytometry. For B and D, asterisks indicate statistical significance (t test; *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001) for comparisons of control and knockdown cells under the same treatment conditions.