Figure 6: In vivo efficacy of a peptide-based FOXM1 inhibitor in patient-derived ALL cells.
From: Identification of FOXM1 as a therapeutic target in B-cell lineage acute lymphoblastic leukaemia
(a) Analysis of FOXM1 target genes after 24 h of 10 μM ARF peptide treatment of patient-derived ALL samples (shown for LAX2), COX6B was used as a reference gene, n=3; ±s.e.m., Student’s t-test was used for statistical analysis. (b) Efficacy of 30 μM ARF peptide on Foxm1+/+ and Foxm1−/− cells after 72 h to confirm specificity; measured by CCK-8 viability assay; n=3; ±s.e.m., Student’s t-test was used for statistical analysis. (c) Confocal microscopy of control or ARF peptide-treated ALL samples (shown for LAX7R), stained for the nucleoli marker fibrillarin (green) and FOXM1 (red) and 4′,6-diamidino-2-phenylindole (DAPI). Scale bar represents 8 μm; representative result of three independent experiments. (d) FOXM1 protein expression after ARF peptide treatment after 24 h (shown for LAX7R) (e) Dose–response for ARF26–44 peptide of pre-B ALL versus B-NHL (green) and CML cell lines (black), ICNI and PDX2 are Ph+ALL, SFO5, LAX7 and LAX7R are normal karyotype ALL (red). (f) Annexin V/DAPI staining after 48 h of 10 μM imatinib in the presence of control peptide or ARF peptide of ICN1. (g) Intracellular ROS levels determined by DCFDA staining in ALL cells treated for 4 h with 10 μM ARF peptide or control peptide (PDX2). (h) Leukaemia burden in mice after injection of 500,000 human luciferase-labelled pre-B ALL cells (LAX7R) and treatment for 10 days with ARF peptide, measured by luciferase bioimaging. (i) A Kaplan–Meier analysis compared overall survival of transplant recipients in the two groups; n=7 per group. Statistical analysis was performed by Log-rank Mantle–Cox test.
