In vitro combination experiments

For in-vitro study, freshly isolated peripheral blood samples were obtained from CLL patients who were not on duvelisib clinical trial (n=5). CLL cells were seeded at 1 × 10⁶ cells/well and treated with either DMSO, 10 μg/mL IgM or a cytokine cocktail containing 1μg/mL sCD40L, 10 ng/mL IL-10, and IL-2 for 4 days⁸. Our previous study in CLL cells has demonstrated activation of BCR pathway through these stimulation and inhibition of this activation by duvelisib.⁸ Cells were either untreated or treated with duvelisib for 4 days. Each culture was treated with or without 0.5–10 nM ABT-199 for 12–16 hours and apoptosis was measured. Cell viability was measured through Annexin V/PI assay.²⁴