Paracrine inhibition of STAT5 phosphorylation in CMV-MoDCs. Monocytes were differentiated into DCs by culture with (MoDC) or without GM-CSF (ΔGM-MoDC) and IL-13. (A) The percentage of MoDCs (Med) and ΔGM-MoDC (ΔGM) cells bearing CD1a and CD83 was determined by flow cytometry with PE-Abs. (B) Total and phosphorylated STAT5 (pSTAT5) analyzed by Western blotting of cell lysates recovered from MoDCs (Med), ΔGM-MoDCs (ΔGM), cells issued from monocytes infected with live CMV (CMV), or UV-inactivated CMV (UVCMV), or monocytes incubated with supernatants from MoDC (SN-MoDC), CMV-MoDC (SN-CMV-MoDC), and UVCMV-MoDC (SN-UVCMV-MoDC). β-actin was used as control. The histograms below represent the quantification of 3 independent experiments. (C) The percentage of cells containing pSTAT5 after infection with BAC-TB40E/GFP was determined by flow cytometry after sequential labeling with a mouse monoclonal Ab specific for pSTAT5 (P-Y694) and PE-labeled anti–mouse Ab. (D) Total and phosphorylated STAT5 proteins were assayed by Western blotting of MoDCs (Med) and CMV-MoDCs (CMV) treated (+) or not (−) with PAA (250 μg/mL) during differentiation. The histogram (right) represents the quantification of 3 independent experiments. Representative of ≥ 3 independent experiments (A,C).