Fig. 2

Transplantation of photoactivated ASCs decreases the accumulation of adipose tissue macrophages. A, B Quantitation by flow cytometry of the proportion of CMFDA+ ATMs in the SVF of EAT from HFD mice that received light-treated ASCs+hydrogel, non-light-treated ASCs+hydrogel, or hydrogel alone. n = 6–8 per group; *p < 0.05 vs. non-light-treated ASCs+hydrogel and hydrogel alone; and ^#p < 0.05 vs. hydrogel alone. Data are mean ± SEM. C–F Immunohistochemical detection of Mac3 (D), CD11c, and CD206 in EAT of from HFD mice that received light-treated ASCs+hydrogel, non-light-treated ASCs+hydrogel, or hydrogel alone. n = 6–8 per group; *p < 0.05 vs. non-light-treated ASCs+hydrogel and hydrogel alone; and ^#p < 0.05 vs. hydrogel alone. Data are mean ± SEM. Macrophages are stained brown, indicated as the ratio of Mac3-positive cells to total cells. Quantification of the pixel intensity of CD11c (E) and CD206 (F) indicating the degree of pro- or anti-inflammatory polarization of macrophage in EAT. Magnification, × 200. Scale bars, 50 μm. a.u.: arbitrary unites. G, H Quantitative RT-PCR analysis of total RNA isolated from EAT of HFD recipient mice for IL-6, IL-1β, TNF-α, MCP-1, CD11c, IL-10, YM1, TNF-β, and CD206 mRNAs. Data were normalized by the amount of 18s mRNA and expressed relative to the corresponding hydrogel alone. n = 6–8 per group. *p < 0.05 vs. non-light-treated ASCs+hydrogel and hydrogel alone; and ^#p< 0.05 vs. hydrogel alone. Data are mean ± SEM. I Serum IL-6 level was measured by ELISA (n = 6/each group); *p < 0.05 vs. non-light-treated ASCs+hydrogel and hydrogel alone; and ^#p > 0.05 vs. hydrogel alone. J The isolated HFD-C57/6J mice-derived peritoneal macrophages were added to upper chambers. ASCs from either light-treated or no-light-treated were placed in the lower chambers. Cells were fixed after 8 h and stained with crystal violet. Migrated cells were counted. Representative images of cell migration are shown. K Quantitative assessment of triplicate cell migration experiments was performed. Data shown are mean ± SEM. *p < 0.05 vs. control (medium without ASCs) and non-light-treated; ^#p < 0.05 vs. control