pHrodo erythrophagocytosis assay using naïve PECs.

<p>A (left panel): CD11b versus F4/80 profile following gating on CD45+ cells allows identifying two distinct populations; (i) CD11b⁺F4/80⁺ cells which represent macrophages within the PEC population and (ii) cells which are negative or low for CD11b and F4/80 expression referred by as the Rest fraction. A (right panel): histogram showing the PE signal obtained following gating on CD11b⁺ F4/80⁺ cells when PECs are incubated alone (black line), with unlabeled RBCs (blue line), with pHrodo-labeled RBCs (orange line) or stimulated with LPS and incubated with pHrodo-labeled RBCs (green line). B: Histogram showing delta medium fluorescence intensity (ΔMFI) obtained by subtracting the PE signal for cells incubated with unlabeled RBCs from cells incubated with pHrodo-labeled RBCs. PECS were unstimulated (white bars) or stimulated with 1μg/ml LPS (black bars). Results are presented +/- SEM and are representative of 4 independent experiments (for each point triplicates were used). (*: p-values ≤ 0.05) C: Microscopic pictures from PECs incubated with pHrodo-labeled RBCs.