The Gene Expression Response of Chronic Lymphocytic Leukemia Cells to IL-4 Is Specific, Depends on ZAP-70 Status and Is Differentially Affected by an NFκB Inhibitor
Figure 5
Effect of an NFκB inhibitor on apoptosis and gene expression response to IL-4 in CLL.
(A) Apoptosis of NBC and CLL cells cultured for 18 h in the absence or presence of IL-4, and CLL cells cultured with IL-4 plus an NFκB activation inhibitor (NFκBi) at 1 µM and 10 µM. CLL are represented together and also separated in ZAP-70 positive and negative. Apoptosis was measured as the percentage of cells labelled with Annexin V (top panel). The bottom panel represents results in CLL patients after subtracting the percentage of apoptotic cells of the Ctrl samples to the IL-4, IL-4 plus 1 µM NFκBi, and IL-4 plus 10 µM NFκBi samples. T tests were used to compare the levels of apoptosis between cell types and conditions. When differences were significant the p-values are indicated as follows: *, p<0.05; **, p<0.01; ***, p<0.001. (B) Expression of the ZAP-70Pos IL-4 targets SUSD2 and CCR2, and (C) of the ZAP-70Neg IL-4 targets AUH, LY75, NFIL3, and BCL6), measured by qPCR, in 7 ZAP-70 positive and 8 ZAP-70 negative patients. The ratios for expression of IL-4 targets following treatment with IL-4 alone, IL-4 plus NFκBi at 1 µM, and IL-4 plus NFκBi at 10 µM, compared to Ctrl, are represented. P-values are depicted as in A.