Fig. 3: In vivo immune cells recruitment and the antigen release and reflux.

A The infiltration of different types of immune cells in hydrogels (CD45⁺ leukocyte, n = 6 mice in each group; F4/80⁺ macrophages, n = 6 mice in each group; CD11c⁺ DCs, n = 6 mice in each group; CD3⁺ T cells, n = 5 mice in each group; CD19⁺ B cells, n = 5 mice in each group; NK1.1⁺ NK cells, n = 5 mice in each group). The experiments were repeated at least two times and representative data from one of the experiments are shown. B The infiltration of different types of immune cells in skins surrounding hydrogels (n = 5 mice in each group). C Staining images of L-Gel (top) and D-Gel (bottom) after subcutaneous injection hydrogels into mice for 1 week. The inflammatory response to the hydrogels were evaluated by a hematoxylin-eosin (H&E) stain, scale bar: 1 mm. Leukocytes, Macrophages, DCs, and T cells were labeled by immunofluorescent biomarker (CD45, F4/80, CD11c, and CD3), scale bar: 100 µm. Nuclei were stained by DAPI to show blue fluorescence. The experiments were repeated at least two times and representative data from one of the experiments are shown. D Quantification of the fluorescence intensity of OVA-Cy5 in injection sites by IVIS Lumina LT (n = 3 mice in each group). The experiments were repeated two times and representative data from one of the experiments are shown. E The LNs reflux of OVA-Cy5 at different time points (n = 5 mice in each group). The experiments were repeated two times and representative data from one of the experiments are shown. Data are presented as the mean ± SEM. Statistical significance was analyzed by two-way ANOVA using Sidak’s posttest (A, B, E). Statistical significance was analyzed by one-way ANOVA using Tukey’s posttest for five groups of data at the same time point and the P value marks the significance difference between L-Gel OVA group and D-Gel OVA group (D). Source data are provided as a Source Data file.