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. 2022 Jul 8;119(28):e2122840119. doi: 10.1073/pnas.2122840119

Fig. 1.

Fig. 1.

Low GGT1 expression promotes the proliferation of ChRCC cells in vivo and in vitro by maintaining high GSH levels. (A) GGT1 protein levels in UOK276 cells expressing EGFP (control) or GGT1. (B) Tumor volume of EGFP- and GGT1-expressing UOK276 xenografts, n = 6 to 7 mice/group. (C) Survival curve for mice in B (n = 6 to 7 mice/group). (D) Proliferation (crystal violet) of UOK276 cells expressing EGFP or GGT1, with and without H2O2 (100 µM) treatment for 24, 48, and 72 h. (E) Proliferation (crystal violet) of UOK276 cells expressing EGFP or GGT1 treated with H2O2 (100 µM) or NAC (1 mM) alone or in combination for 48 h. (F) Proliferation (crystal violet) of UOK276 cells expressing EGFP or GGT1 treated with TBH (15 µM) or NAC (1 mM) alone or in combination for 48 h. (G) Volcano plot of differential metabolites in UOK276 cells with GGT1 expression compared with EGFP control (n = 4 biological replicates per condition). Blue signifies downregulation in GGT1-expressing cells, Red signifies upregulation. (H) GSH concentration in UOK276 cells with EGFP or GGT1 expression (n = 3 biological replicates per condition). (I) ROS level (Deep Red assay) in UOK276 cells with EGFP or GGT1 expression and with NAC (1 mM, 24 h) (n = 3 biological replicates per condition). (J) GSH concentration in EGFP- and GGT1-expressing UOK276 xenograft tumors shown in B (n = 4 biological replicates per condition). (K) Cystine-FITC (5 µM, 3 h) uptake in UOK276 cells with or without GGT1 expression, with or without IKE (1 µM, 3 h) treatment (n = 3 biological replicates per condition). Error bars represent mean ± SD. Statistical analyses were performed using two-tailed Student’s t test or one-way ANOVA if more than two groups. *P < 0.05; **P < 0.01; ****P < 0.0001.