Fig. 5.

Cinobufotalin increased the sensitivity of FOXO1-overexpressing NPC cells to DDP. a Dose–response curves are shown for HONE1-EBV + -FOXO1 and 5–8F-FOXO1 after treatment with DDP and/or CB for 48 h. b Survival analysis data show the cumulative overall survival time of mice in the FOXO1 + DDP group, FOXO1 + CB-treated group, FOXO1 + DDP + CB group and control group (n = 10, log-rank test). c Mean weight of mice in the FOXO1 + DDP group, FOXO1 + CB-treated group, FOXO1 + DDP + CB group and control group (n = 10, log-rank test) are shown. Sphere-forming assay (d, scale bar: 500 μm), migration assay, invasion assays (e, scale bar: 200 μm) and scratch assays (f, scale bar: 200 μm) of NPC cells were performed after transfection with the FOXO1 lentiviral vector and CB (750 nM) as indicated. Student’s t-test. Mean ± s.d. *P < 0.05; **P < 0.01; ***P < 0.001. g Expression levels of MYH9, PI3K, p-PI3K, AKT, p-AKT, C-Myc, P53, β-catenin, GSK3β, p-GSK3β, and ubiquitin in FOXO1-suppressed NPC cells treated with CB. h MYH9 expression was measured in FOXO1-suppressed NPC cells treated with CB; data are normalized to ARF. One-way ANOVA and Dunnett multiple comparison tests were used for analysis. **P < 0.01 ***P < 0.001. i miR-133a-3p expression was measured in FOXO1-suppressed NPC cells treated with CB; data are normalized to U6. One-way ANOVA and Dunnett multiple comparison test. *P < 0.05 ***P < 0.001. j Expression levels of PI3K, p-PI3K, AKT, p-AKT, C-Myc, P53, and MYH9 in FOXO1-suppressed NPC cells treated with CB. k Amplification of mir-133a-3p-binding sites after Ch-IP using an antibody against P53 in FOXO1-suppressed NPC cells treated with CB. An IgG antibody was used as the negative control. Student’s t-test. Mean ± s.d. *P < 0.05; **P < 0.01. l Expression of N-Ca, E-Ca and Vimentin, OCT4, SOX2, and β-actin following CB treatment in FOXO1-suppressed NPC cells