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Fig. 1 | Journal of Experimental & Clinical Cancer Research

Fig. 1

From: Pharmacological modulation of Kv1.3 potassium channel selectively triggers pathological B lymphocyte apoptosis in vivo in a genetic CLL model

Fig. 1

Kv1.3 expression and effect of PAPTP on pathological human and murine lymphocytes ex vivo. a Kv1.3 expression was assessed by flow cytometry on ex vivo CLL B cells isolated from 7 CLL patients and treated for 24 h with 1 μM PAPTP. Kv1.3 expression, reported as mean fluorescence intensity (MFI, arbitrary units), is represented as a function of the viability of CLL B cells, estimated as the percentage of living cells (compared to non-treated ones) after treatment. The statistical relationship is assessed with Pearson’s correlation coefficient. Inset: Chemical structure of PAPTP. b Mitochondrial ROS production and mitochondrial membrane potential were measured, respectively, by MITOSOX (green lines) and TMRM (purple lines) fluorescence from ex vivo CLL B cells, isolated from 6 CLL patients, treated with 1 μM PAPTP or left untreated. The percentage of the initial values of MITOSOX and TMRM signals are reported as a function of treatment time and are shown as mean ± SD. Statistical significance of differences was assessed with the Multiple unpaired t test. Inset: Cytochrome c levels in the cytoplasm were evaluated by western blotting from ex vivo human CLL B cells treated with DMSO or PAPTP 1 μM (25 μg protein/lane). β-actin was chosen as cytoplasmic marker and is shown as well. c CLL B cells mortality was assessed after 24 h. Leukemic cells from 6 patients were cultured with allogeneic mesenchymal stromal cells (MSCs) as in [7] and treated with DMSO or PAPTP 1 μM. Data are shown as mean ± SEM. Statistical significance of differences was assessed with the Wilcoxon test. d Kv1.3 expression in CD19+CD5− and CD19+CD5+ cells from peripheral blood of Eμ-TCL1 mice was assessed by flow cytometry. Kv1.3 expression is reported as mean fluorescence intensity (MFI, arbitrary units). Data were obtained from 31 individual animals and are shown as mean ± SEM. Statistical significance of differences was assessed with the Wilcoxon test. e A representative fluorescein isothiocyanate (FITC) fluorescence distribution of the populations shown in panel d. f Percentage of CD19+ dead cells, estimated with Annexin V staining, was assessed by flow cytometry on ex vivo blood cells (isolated from WT and Eμ-TCL1 mice), treated with DMSO or 5 μM PAPTP. Data were obtained from 4 independent experiments and are shown as mean ± SEM. Statistical significance of differences was assessed with the Mann–Whitney test

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