Expression of FMNL1 in normal and malignant tissue. (A) Relative quantitative mRNA expression of FMNL1 in different tissues pooled from different healthy donors was measured using quantitative real-time PCR. The relative quantitative expression compared with skeletal muscle was calculated using the delta-delta Ct method. (B) Relative quantitative mRNA expression of FMNL1 in stimulated PBMCs and PBMC-derived subpopulations is shown. Subpopulations were isolated by negative (CD4, CD8, CD14, CD19) and positive (CD34) magnetic bead depletion. Monocyte-derived dendritic cells (DCs) were isolated by adhesion, cultured in presence of IL-4 and GM-CSF, and matured with IL1β, IL-6, TNF-α, and prostaglandin E2. For activation of PBMCs, cells were incubated for 3 days using IL-2 and OKT3; B-cell activation was induced by incubation with CD40L-transfected NIH3T3 cells for at least 3 weeks. Error bars indicate the standard deviation of different samples tested. (C) Relative quantitative mRNA expression of FMNL1 in CLL samples and tumor cells derived from patients with acute B- and T-ALL as well as AML is shown in comparison with normal PBMCs. (D) FMNL1 protein expression in PBMC-derived cells, lung fibroblasts, and nontransfected 293 HEK cells by Western blot analysis. Total protein (50 μg) was used for sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE). 293 HEK cells transfected with FMNL1 were used as positive control. Probing with antiactin antibody served as loading control. (E) FMNL1 protein expression in native tumor cells and transformed cell lines by Western blot analysis. Total protein of native leukemic cells (50 μg) and total protein of lysed cell lines (100 μg) was used for SDS-PAGE. 293 HEK cells transfected with FMNL1 were used as positive control. Probing with antiactin antibody served as loading control.