Fig. 3

Chronic exposure to TNFα leads to eEPC-to- macrophage conversion. A Representative photographs showing the morphological change of eEPCs in response to chronic TNFα exposure. Cells were incubated with TNFα at 50 ng/ml for 6 days. B Characterization of eEPCs. eEPCs were subjected to fluorescence staining with Dil-ac-LDL (red) and lectin (green). Cell nuclei were visualized by DAPI (blue). Dual positives were counted (n = 3). C Characterization of macrophage lineage. eEPCs were subjected to fluorescence staining with anti-KUL-01 (green). The cell nuclei were labeled with DAPI (blue). MRC1+ cells were counted (n = 6). D eEPCs were exposed to TNFα at 50 ng/ml for 3–6 days and subjected to immunofluorescence analysis with anti-CD133 (green). The cell nuclei were labeled with DAPI (blue). CD133+ cells were counted (n = 6). E Phagocytosis capability assay. eEPCs were exposed to TNFα at 50 ng/ml for 3–6 days and allowed to phagocyte E. coli (green). The cell nuclei were labeled with DAPI (blue). Chicken PBMNC-derived macrophages were used as a positive control. Cells having intracellular bacteria were counted (n = 6). The data are representative of 2 separate experiments. Photographs are from one representative experiment. Data were expressed as median ± 95% confidence interval (Mann–Whitney). *P < 0.05. **P < 0.01