FIG. 3.
M2r macrophages retain an M2 signature after secondary proinflammatory stimulation both in vitro and in vivo. A: Schematic experimental set-up. Primary activation with M0-, M2r-, or M1-inducing stimuli was for 24 h, and secondary activation for an additional 24 h with LPS/IFNγ (48 h). B: IL-10 production assessed by ELISA in secondary-activated macrophages. C: Flow cytometry analyses of CD86 and PD-L2 expression on primary- and secondary-activated macrophages. Histograms and mean fluorescence intensity analyses are depicted. MFI, mean fluorescence intensity. D: Selected gene expression assessed by RT-PCR after primary and secondary activations of macrophages. E: Ex vivo analyses of CD86 and PD-L2 on CD11b+DiD+ macrophages recovered from pancreata (left) or PLNs (right). Histograms and percentage of cells depicted. The results are representative of two independent experiments (n = 4). *P < 0.05.