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. 2023 Jan 13;42(4):e110620. doi: 10.15252/embj.2022110620

Figure 7. AGI6780 restores chemosensitivity in chemo‐resistant UC cells.

Figure 7

  1. Graph showing the viability of GR cells exposed to various concentrations of GEM for 48 h in combination with DMSO or 10 μM AGI6780 (upper: T24GR, lower: UMUC3GR). The data are shown as the mean values ± SDs (n = 3, biological replicates).
  2. Relative 2‐HG production level based on targeted CE‐MS metabolomics in GR cells exposed to DMSO or 10 μM AGI6780 for 24 h (upper: T24GR, lower: UMUC3GR). The data are shown as the mean values ± SDs (n = 3, biological replicates) and were analyzed by Student's t‐test. *P < 0.05, **P < 0.01.
  3. Relative dCTP level based on targeted CE‐MS metabolomics in GR cells exposed to DMSO or 10 μM AGI6780 for 24 h (upper: T24GR, lower: UMUC3GR). The data are shown as the mean values ± SDs (n = 3, biological replicates) and were analyzed by Student's t‐test. **P < 0.01, ***P < 0.001.
  4. Graph showing the viability of GR cells exposed to various concentrations of CDDP for 48 h in combination with DMSO or 10 μM AGI6780 (upper: T24GR, lower: UMUC3GR). The data are shown as the mean values ± SDs (n = 3, biological replicates).
  5. NADPH/NADP ratio in GR cells exposed to DMSO or 10 μM AGI6780 for 24 h. The data are shown as the mean values ± SDs (n = 3, biological replicates). The data were analyzed by Student's t‐test (upper: T24GR, lower: UMUC3GR). *P < 0.05, **P < 0.01.
  6. Intracellular ROS generation in GR cells exposed to DMSO or 10 μM AGI6780 for 24 h. The data are shown as the mean values ± SDs (n = 3, biological replicates). The data were analyzed by Student's t‐test (upper: T24GR, lower: UMUC3GR). *P < 0.05, **P < 0.01.
  7. In vivo treatment of orthotopically implanted mouse models (n = 5 mice per group). UMUC3GR cells (2 × 106 cells) were implanted subcutaneously into the flanks of nude mice. One week after implantation, the groups were treated with GEM alone (i.p. injection of 50 mg/kg weekly), AGI6780 alone (i.p. injection of 50 mg/kg daily), or a combination of GEM and AGI6780 (i.p. injection of 50 mg/kg GEM weekly and i.p. injection of 50 mg/kg AGI6780 daily). The mean tumor volume (mm3) ± SD for each group is shown. Data were analyzed by Student's t‐test. **P < 0.01.
  8. Representative images of tumors upon necropsy.
  9. Body weights of mice with the indicated treatments. The data are shown as the mean values ± SDs (n = 5 mice per group), and were analyzed by Student's t‐test. n.s. = non‐significant.
  10. IHC staining for IDH2, Ki‐67, CAIX, TIGAR, TKT, and CTPS1 in groups with GEM alone, AGI6780 alone, and the combination of GEM and AGI6780.
  11. In vivo treatment of orthotopically implanted mice (n = 5 mice per group). UMUC3GR cells (2 × 106 cells) were implanted subcutaneously into the flanks of nude mice. One week after implantation, the groups were treated with CDDP alone (weekly i.p. injection of 15 mg/kg), AGI6780 alone (daily i.p. injection of 50 mg/kg), or a combination of GEM and AGI6780 (weekly i.p. injection of 15 mg/kg CDDP and daily i.p. injection of 50 mg/kg AGI6780). The tumor volume (mm3, mean ± SD) in for each group is shown. Data were analyzed by Student's t‐test. **P < 0.01.
  12. Representative images of tumors upon necropsy.
  13. Body weights of mice treated as indicated. The data are shown as the mean values ± SDs (n = 5 mice per group) and were analyzed by Student's t‐test. n.s. = non‐significant.

Source data are available online for this figure.