Figure 3.

ILC2s protect liver from IRI by increasing the proportion of M2 CD45+CD11b+F4/80high macrophages (A) Representative FACS analysis showing that exogenous IL-33 administration significantly increased the proportion of ILC2s in liver as compared with PBS controls and IL-33 receiving anti-CD90.2 administration, which significantly depleted ILC2s. (B) Sketch map of medicine administration and IRI surgery. Mice were treated with exogenous IL-33 daily for 5 consecutive days as well as anti-CD90.2 antibody twice before IRI surgery. Mice were euthanized 12 h after reperfusion. (C) HE staining of Sham, PBS, IL-33 and IL33&anti-CD90.2 groups as determined at 12 h after reperfusion (×200). (D) Suzuki’s Scores resulting from IR-induced liver injury in Sham, PBS, IL-33 and IL33&anti-CD90.2 groups (n=6 per group). (E) Representative cell apoptosis immunofluorescence of the 4 groups as determined at 12 h post-reperfusion. Cell apoptosis was measured using TUNEL (red) and Caspase3 (green). Apoptotic cells display a red nucleus (TUNEL) and green cytoplasm (Caspase-3) while normal cells show blue nuclei (DAPI) (×400). (F) Histograms of quantitative analysis of TUNEL-positive cells (n = 6 per group). (G) Representative FACS analysis showing the proportion change of M2 CD45+CD11b+F4/80high macrophages. (An CD206+ increase in proportion was assumed as a M2 polarization among CD45+CD11b+ F4/80high cells) (H) Quantitative analysis of percent of M2 CD45+CD11b+F4/80high macrophages (n = 5 per group). (I) Relative gene expressions of IL-1β, iNOS, HO-1, FIZZ1 and IL-10 in macrophages purified from mice liver in different groups. (***P<0.05, ***P<0.001).