Fig. 6: Macrophage GRIP1 contributes to phagocytic activity of M2_IL4 and M2_GC in vitro and tissue healing in vivo.

a Relative fluorescence intensity of WT and GRIP1-cKO M2_IL4, M2_Cort, and M2_Dex relative to that in WT M0 (set to 100%) after exposure to pHrodo Green S. aureus BioParticles (see “Methods”). Shown are mean ± SEM (n = 3). b Histopathological changes assessed by H&E staining of colons from WT and cKO mice 2 wks after initiating DSS-induced colitis. Grades 1, 2, and 3 are defined in Methods. Scale bar = 20 μm. Plotted is the total area representing grade 3 for colon specimens of WT and GRIP1-cKO. Shown are mean ± SEM (n = 4). c The expression of indicated genes in total RNA from colons of WT and cKO mice was measured by RT-qPCR. The expression of each gene was normalized to β-actin, and then the fold difference to baseline, non-treated (calibrant) value was calculated for WT or cKO. Shown are mean ± SEM (WT n = 7 (Il23) 10 (Mrc1), 11 (Cd163), 12 (Il10, Tnf), 13 (Klf4, Arg1, Il1b), 14 (Pparg) or 15 (Grip1, Klf9); cKO n = 5 (Il23), 6 (Mrc1, Il10), 7 (Pparg, Arg1), 8 (the rest)). d, e Cells were isolated from the large intestine lamina propria at day 11, and single-cell suspensions incubated in FACS buffer with conjugated antibodies to CD45-PE, CD11b-PE-Cy7, Ly6C-PerCP-Cy5.5, Ly6G-APC, and CD206-AF700. Dead cells were excluded using DAPI. Cell sorting was performed on Symphony S6 (BD Biosciences; see Supplementary Fig. 6d for FACS strategy), and (d) data analyzed with FlowJo v10.10 software. Ly6C⁺, CD206⁺, and Ly6C⁺CD206⁺ cells were pooled, total RNA was isolated, and expression of indicated genes was assessed by RT-qPCR (e). Shown are mean ± SEM (n = 4). P-values are indicated; ns, non-significant (p ≥ 0.05). In a–e, n = the number of independent biological replicates (mice), and significance is determined by a one-tailed unpaired t test. Source data are provided as a Source data file.