Figure 1: IL-4 produced by AC2M2 cells promotes M2-like macrophage activation but does not promote autocrine proliferation effects. A: Conditioned medium (CM) collected from EV-AC2M2 (EV CM) or IL4-AC2M2 (IL-4 CM) cells was added to BMA cell cultures at increasing CM concentrations of 5%, 20%, 50% or 100%, and cultured for 24 hours. BMA cells were also challenged with 2.5, 5 or 10 ng/ml recombinant mouse IL-4 (rIL-4) for positive controls. Lysates were analyzed by immunoblotting for macrophage polarization using anti-arginase I (Arg1) antibody, and Jak/Stat activation using anti-phospho-Jak1 (pJak1), anti-phospho- Jak3 (pJak3) and anti-phospho-Stat6 (pStat6) antibodies. B: EVAC2M2 or IL4-AC2M2 cells were seeded in triplicate (n=3) on 6- well plates. Four hours later or at the indicated days post plating, cells were trypsinized, and cell numbers were counted using Z1 Coulter Particle Counter. No significant difference was observed in growth rates in vitro between EV-AC2M2 and IL4-AC2M2 cells (P=0.3654).