CAL-101 antagonizes alternative microenvironment stimuli activated by PI3K pathway. (A) CD19⁺ cells from patients with CLL (n = 5-10) were incubated with or without various doses of CAL-101 and 50 ng/mL BAFF for 48 hours. (B) CD19⁺ cells from patients with CLL (n = 5) were incubated with or without various doses of CAL-101 and 20ng/mL TNF-α for 48 hours. (C) CD19⁺ cells from patients with CLL (n = 5-10) were incubated with or without various doses of CAL-101 on and off fibronectin-coated plates for 48 hours. (D) CD19⁺ cells from patients with CLL (n = 7) were isolated from peripheral blood and incubated with or without 1 or 10μM CAL-101 in suspension or on an HS-5 cell layer for 48 hours. Viability was determined by annexin/PI flow cytometry and is shown relative to time-matched untreated controls for each group. Horizontal lines represent the mean. (E) CD19⁺ cells from patients with CLL (n = 4) were incubated with 1 μg/mL CD40L, 50 ng/mL BAFF, and 20 ng/mL TNF-α for 2 hours. AKT phosphorylation at Ser473 was assessed by immunoblot. Results are shown from 1 of 4 experiments.