Fig 1: Inducible expression of CD127 in monocyte-derived macrophages. (A) Selected immune cells from the innate and adaptive immune system were isolated from Buffy Coats from healthy donors. Some cell types (Neutrophils, CD14+ monocytes, and naive CD4+ T cells were lysed directly after the isolation. Some CD14+ monocytes were matured into Monocyte-derived macrophages, immature and mature DCs as described in the methods section. The amount of CD127 and CD132 was analyzed via Western Blot with β-Actin as reference. One out of two experiments (n=2) is shown as demonstration. (B) mRNA levels of CD127 and CD132 relative to housekeeping GAPDH mRNA in freshly isolated monocytes and DCs, which were stimulated with 10 ng/ml lipopolysaccharide (LPS) for the displayed time points. Standard deviations of two experiments (n=2) performed with three replicates each are shown. *P < 0.05 for comparison of the indicated time points with baseline. (C) mRNA levels of CD127 and CD132 relative to housekeeping GAPDH mRNA in monocyte-derived macrophages, which were stimulated with 10 ng/ml LPS, 100 ng/ml poly(I:C) HMW or 100 ng/ml pams3CSK4, with or without 5 µg/ml antibody against IFNAR1, for the time points shown. Standard deviations of three experiments (n=3) performed with three replicates each are shown. *P < 0.05 for comparison of the indicated time points with baseline. (D) Monocyte-derived macrophages were stimulated with 10 ng/ml LPS for the displayed time points. The amount of CD127 and CD132 was assessed by Western Blot (top panel) and by immunofluorescence (bottom panel). One out of two experiments (n=2) is shown as demonstration.