Fig. 6: N-8 attenuates the progression of AML in vivo.

A Schematic strategy for investigating the effect of N-8 on the progression of AML in mice xenografted with U937 AML cells (AML mice). Ara-C, cytarabine. po, Per Os. ip, Intraperitoneal. QD, Quaque Die. QOD, Quaque Other Die. B Flow cytometry analysis of the percentage of leukemia cells in the PB, BM, and spleen of the indicated groups (n = 5 mice per group). hCD45⁺ cells were quantified using FlowJo software. Data are presented as the mean ± S.E.M. Statistical significance was calculated using a one-way ANOVA. C Flow cytometry analysis of the percentage of apoptotic cells in the PB, BM, and spleen of the indicated groups (n = 5 mice per group). Annexin V⁺ cells were quantified using FlowJo software. Data are presented as the mean ± S.E.M. Statistical significance was calculated using a one-way ANOVA. D The protein levels of BCL2, BCL-XL, and MCL1 in the spleen of AML mice treated with or without N-8 were detected using western blotting. E Kaplan-Meier survival curves for the indicated mice (n = 5 mice per group). Statistical significance was calculated using a two-sided log-rank test. F The body weight curves of AML mice treated with the indicated agents (n = 5 mice per group). Data are presented as the mean ± S.E.M. Statistical significance was calculated using a two-way ANOVA. G Schematic of the strategy used to investigate the effect of N-8 on the progression of AML in the MLL-AF9 leukemia model. VEN, venetoclax. po, Per Os, QD, Quaque Die. H Representative spleen images (left) and statistical analysis of spleen weights (right) obtained from mice with the indicated treatment. Scale bar, 1 cm. I Flow cytometry analysis of the percentage of leukemia cells in the PB, BM, and spleen of the indicated groups (n = 5 mice per group). GFP⁺ cells were quantified using FlowJo software. Data are presented as the mean ± S.E.M. Statistical significance was calculated using a one-way ANOVA. J Flow cytometry analysis of the percentage of apoptotic cells in the PB, BM, and spleen of the indicated groups (n = 5 mice per group). Annexin V⁺ cells were quantified using FlowJo software. Data are presented as the mean ± S.E.M. Statistical significance was calculated using a one-way ANOVA. K The protein levels of BCL2, BCL-XL, and MCL1 in the spleen and BM of AML mice treated with or without N-8 was detected using western blotting. L Kaplan-Meier survival curves for the indicated mice (n = 5 mice per group). Statistical significance was calculated using a two-sided log-rank test. M The body weight curves of leukemia mice treated with the indicated agents (n = 5 mice per group). Data are presented as the mean ± S.E.M. Statistical significance was calculated using a two-way ANOVA. N Schematic diagram illustrating the mechanism by which CTSD inhibition induces cell apoptosis.