Effect of combined treatment with AQX-435 and ibrutinib on anti-IgM-induced AKT phosphorylation. A and B, CLL samples were pre-treated with AQX-435 (30 μM) and/or ibrutinib (100 nM), or DMSO as a control, for 30 minutes. Cells were then stimulated with bead-bound anti-IgM or control antibody for 15 minutes and expression of total/ phospho-AKT (S473) was analyzed by immunoblotting. HSC70 was also analyzed as an additional loading control. A, Representative results and B, quantitation for all samples analyzed (n=8). Graph shows mean (±SD) AKT phosphorylation relative to control (set to 1.0) and the statistical significance of the indicated differences (Student’s t-test).