Intracellular TNF-α and IL-4 expression in gated CD33+ monocytes by three-color flow cytometry. Intracellular TNF-α and IL-4 expression in gated CD33+

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Intracellular TNF-α and IL-4 expression in gated CD33+ monocytes by three-color flow cytometry. Intracellular TNF-α and IL-4 expression in gated CD33+ monocytes by three-color flow cytometry. PBMCs cultured for 6 h in medium (left set of panels) or stimulated with PMA-ionomycin (middle set of panels) or PHA (right set of panels) in the presence of monensin were labeled for surface expression of CD33 (PE/Cy5) and CD14 (FITC) and for intracellular TNF-α or IL-4 by using PE-conjugated MAbs. CD33+ gated monocytes were analyzed on FL1 (FITC) versus FL2 (PE) dot plots to discriminate cytokine expression in CD14− and CD14+ monocytes. Data from one representative experiment of five experiments performed are shown. Jarołsaw Baran et al. Clin. Diagn. Lab. Immunol. 2001; doi:10.1128/CDLI.8.2.303-313.2001