Figure 2: Regulation of FOXM1 expression in patient-derived pre-B ALL cells.
From: Identification of FOXM1 as a therapeutic target in B-cell lineage acute lymphoblastic leukaemia
(a) Foxm1 expression levels of three matched samples of IL-7-dependent B-cell precursors (in the presence of IL-7) and BCR-ABL1-transformed ALL-like cells (IL-7 independent). (b,c) Immunoblot analysis for the expression of FOXM1 in normal CD19+ B cells (b) derived from peripheral blood (PB) or CD19+ CD10+ B-cell precursors (c) derived from BM and patient-derived pre-B ALL; loading order for ALL: ICN1 (Ph+), PDX2 (Ph+), SFO5 (normal karyotype), LAX7 (normal karyotype), LAX7R (normal karyotype), LAX2 (Ph+) and PDX2 (Ph+), MPX2 (Ph+), MPX3 (Ph+), MPX5 (Ph+), BLQ5 (Ph+), respectively; shown with β-actin as loading control. Mean densitometric values relative to loading control are shown in the bar graphs ±s.e.m., significance was determined by Student’s t-test. (d) Time course of FOXM1 protein levels after TKI treatment (shown for ICN1), β-actin was used as a loading control; representative result of three independent experiments is shown. (e) Human Ph+ ALL cells (PDX2) were treated with TKI at low doses for 96 h and analysed for FOXM1 expression with β-actin as a loading control with and without overexpression of BCL2, representative result of three independent experiments is shown. (f) Time course analysis of FOXM1 expression in PDX2 with either empty vector or BCL2 overexpression. (g) Immunoblot analysis of Foxm1 with and without the expression of constitutively active Myr-Akt in murine ALL, treated with low concentrations of TKI or control for 96 h. (h) Immunoblot analysis Foxm1 expression in murine BCR-ABL1-transformed Foxo3a+/− and Foxo3a−/− ALL treated with low concentrations of TKI or control for 4 days; representative result of three independent experiments is shown. (i) Foxm1 qRT–PCR was performed on day 2 after transduction with EV or a constitutively active form of FOXO3a (FOXO3aAAA), Hprt was used as a reference, n=3; ±s.e.m., Student’s t-test was used for statistical analysis. (j) Foxm1 protein levels were measured 3 days after the introduction of a constitutively active form of FOXO3a (FOXO3aAAA), with β-actin as a loading control.
