Fig. 2: Evaluation of canonical and non-canonical NF-κB activity in del(11q)/BIRC3-deficient CRISPR/Cas9-engineered CLL cells.

A ELISA measurement of relative NF-κB family transcription factor DNA-binding activity in nuclear extracts from HG3-edited clones. Left panel shows DNA-binding activity of NF-κB transcription factors involved in the canonical signaling (p65, c-Rel, and p50). Right panel displays the DNA-binding activity of non-canonical NF-κB transcription factors (p52 and RelB). Data are represented as the mean ± SD. B Whole-cell, cytoplasmic and nuclear lysates of HG3-del(11q) clones analyzed by immunoblotting for NIK, p-IKKα/β, p-NF-κB2, NF-κB2 (p100/p52), NF-κB1 (p105/p50), and RelB proteins. GAPDH was used as loading control for whole-cell and cytoplasmic lysates and H3 was used as loading control for nuclear extracts. Relative quantification for each protein (mean of three clones per condition) is depicted in Supplementary Fig. 3b. C Whole-cell lysates from HG3^WT, HG3-del(11q), and HG3-del(11q) BIRC3^MUT analyzed by immunoblotting for BCL2 family members: BCL2, BCL-xL, MCL1, BIM, NOXA, BAK, and BAX. β-actin was used as loading control. Relative quantification for each protein (mean of three clones per condition) is detailed in Supplementary Fig. 4a.