Fig. 4: Differential modulation of macrophage phagocytosis by GelMA crosslinking. | Nature Communications

Fig. 4: Differential modulation of macrophage phagocytosis by GelMA crosslinking.

From: Hydrogel crosslinking modulates macrophages, fibroblasts, and their communication, during wound healing

Fig. 4

a Dot plot of phagocytosis- related genes expressed by M3 and M4 cells in lo- and hi-GelMA treated wounds. Dot size corresponds to the proportion of cells within the group expressing each transcript, and dot color correspond to the expression level. b Representative laser scanning microscopy images of labeled BMDM (green) seeded on pHrodo-labeled (red) hi- or lo-GelMA for 16h, along with quantification. Cyan arrowheads: phagocytosed GelMA. Scale bar: 25 µm. n = 3 (BMDM harvests from individual mice). p = 0.005 (t-test, two-sided). c Violin plot of integrin gene expression across in vivo Mϕ subpopulations (bottom) and qPCR validation (top) of integrin gene expression in BMDM cultured on GelMA for 24h, compared to BMDM cultured without GelMA (control), n = 3 (BMDM harvests from individual mice). Left to right: p = 0.003, 0.002, 0.006, 0.005, 0.01(t-test, two-sided). d Violin plot of complement components gene expression across in vivo Mϕ subpopulations (bottom) and qPCR validation (top) of complement component gene expression in BMDM cultured on GelMA for 24h, compared to BMDM cultured without GelMA (control). For C3, expression in lo versus hi-GelMA in vivo and in vitro. n = 3 (BMDM harvests from individual mice). C3 compared to no GelMA: p = 0.001; C3 lo/hi: p = 0.007 (t-test, two-sided). e Fluorescence signal of BMDM seeded on pHrodo-labeled (red) hi- or lo-GelMA for 16h, treated with 10 µg/ml blocking antibodies (CD18, CD11c, CD51, CD11a, and CD11b), 10 µM phagocytosis inhibitor Cytochalasin-D (Cytoch.D), or antibody isotype controls (Isotype). n = 3 (BMDM harvests from individual mice). Left to right: p = 0.01, 0.01, 0.004, < 0.0001, < 0.0001 (one-way ANOVA, Dunnett). f Fluorescence signal measurement of BMDM seeded on unlabeled gels stained with lysotracker (green) under conditions similar to (e). Left to right: p = 0.05, 0.05, 0.01, 0.01, 0.001 (one-way ANOVA, Dunnett). For microscopy images of (e) and (f) see Supplementary Fig. 8b. n = 3 (BMDM harvests from individual mice). All data presented as mean ± SD. Source data are provided as a Source Data file.

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