HNP1-3 inhibit M-CSF–induced cytokine production during differentiation. (A) Healthy donor monocytes were incubated with 100 ng/mL rhM-CSF and 5μM rhHNP3 for 1 and 5 days. Cytokines were detected in their culture supernatant using an antibody array. (B) RQ-PCR analysis of indicated cytokine gene expression in monocytes exposed to 100 ng/mL rhM-CSF in the absence or presence of rhHNP3 for 1 and 5 days. (C) HNP1-3 were depleted from CD14⁻/CD24⁺ CMML supernatant using clone 21 antibody coupled to beads as in Figure 5. CD14⁺/CD24⁻ CMML cells were cultured with M-CSF for 2 days in the supernatant (depleted, not depleted, or depleted and supplemented with the 3 rhHNPs; total concentration: 5μM), before assessing indicated gene expression. Mean ± SD of triplicates; 1 of 3 independent experiments is shown.