Fig. 6

Treatment of mice bearing melanoma xenografts with TMZ in combination with AG-120. A Schematic represents the treatment model after 1 × 10⁶ A375 melanoma cells were injected subcutaneously into the flank of nude mice. A separate experiment with B16-F10 melanoma murine cells involved 4 × 10⁴ cells injected subcutaneously into C57BL/6 J recipient mice. After 8-10 days, when tumors reached 100-120 mm³, mice were divided into four groups and treated with i) Vehicle; ii) TMZ (30 mg/kg) every day; iii) AG-120 (150 mg/kg) twice a day; iv) AG-120 + TMZ (150 mg/kg + 30 mg/kg). B Tumor growth curves of A375 melanoma xenografts in nude mice. Tumor sizes were assessed twice per week using calipers (n = 5 per group), C Average tumors weights (mg) of A375 xenografts in each group (n = 5 per group). D Representative image of excised and in vivo tumors of A375. E Body weights of A375 melanoma xenografts in nude mice (n = 5 per group). F Cell proliferation in tumor xenografts was estimated by nuclear immunolabeling (Ki-67). Scale bar, 50 μm. Quantitation is shown below from four random fields per section. G Tumor xenograft apoptosis was estimated with labeled cleaved caspase-3. Quantitation is shown below from four random fields per section. Scale bar, 50 μm. Each data point represents the mean ± SEM of at least three independent experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001