Fig. 7: Analysis of BCR inhibitor sensitivity of NFKBIE-wt and NFKBIE-ko CLL cells.

A Changes in NFKBIE MAF during ibrutinib treatment in vitro. NFKBIE-ko and NFKBIE-wt TCL1-355 TKO cells were mixed at a 1:3 ratio and cultured in the presence or absence of ibrutinib (0.2 or 1.0 µM). DMSO was present at a concentration of 0.5% in both the ibrutinib-treatment and the control condition. DNA was isolated at the indicated time points for MAF analysis. Summary of the data is shown in the left panel and analysis of one representative sample from each condition in the right panels. Statistical analysis was done using One Way ANOVA with Tukey test for multiple comparisons. B Growth curve analysis of NFKBIE-wt and NFKBIE-ko CLL cells in culture with the indicated inhibitors. Each data point represents an average of 3 independent experiments. Graph in right panel shows changes in NFKBIE MAF during R406 and idelalisib treatment in vitro. NFKBIE knockout and wild type TCL1 leukemia cells were mixed as in A and cultured in the presence or absence of 1.0 μM idelalisib or R406. DMSO was used at a concentration of 0.5% in all investigated conditions. Statistical analysis was done using One Way ANOVA with Holm-Sidak test for multiple comparisons. C Changes in NFKBIE MAF during ibrutinib treatment in vivo. NFKBIE-knockout and NFKBIE-wild type leukemia cells were mixed at a 1:3 ratio and injected in 16 wild-type mice. Treatment was started three days later (8 mice treated with ibrutinib and 8 with vehicle control) and lasted for 2 weeks. The MAF at the end of treatment for the control (Ctrl) or ibrutinib (Ibr)-treated group is shown in the graph. The PB and PC samples from 2 ibrutinib-treated mice were lost during processing and were not available for analysis. Statistical analysis was performed with the t test (PC and PB) and Mann-Whitney rank sum test (spleen).