Fig 2: CD63 expression results for two different desensitization approaches in bone-marrow-derived mast cells (BMMCs)(A) CD63 expression in different in-between step points of the desensitization protocol is reduced when the concentration increase between steps is lessen.(B) CD63 expression of sensitized and desensitized (DS) BMMCs after a single dose challenge is lower when the concentration increase between steps is reduced. N = 1 experiment.

Fig 3: Polyclonal desensitization impairs mast cell (MC) degranulation after a single-dose allergen challenge(A) % of β-hexosaminidase activity (left), and CD63 and CD107a expression (middle and left) by flow cytometry of desensitized bone-marrow-derived MCs (BMMCs) after a single-dose peanut (PN) challenge.(B) % of β-hexosaminidase activity (left), CD63 and CD107a expression (middle and left) of desensitized LAD2 after a single-dose anti-hIgE or PN challenge. Comparisons between 2 groups were made with a two-tailed t-student or Mann-Whitney tests. The latter was used when data were non-parametric. Welch correction was used when equal SD between data could not be assumed. Data are shown as mean±SEM. N = 4–8 independent experiments. p<0.0332 (∗); p<0.0021 (∗∗); p<0.0002 (∗∗∗); p<0.00001 (∗∗∗∗).

Fig 4: Characterization of sEVs from ADSC culture medium, which are delivered to kidney and muscle of old mice.(A) Nanoparticle tracking analysis (NTA) of sEVs from young ADSCs. (B) Western blot image of staining with TSG101 and bovine serum albumin, a sample of purified sEVs from the cell culture medium of ADSCs, and a sample of FBS present in the same medium are shown. (C) Flow cytometry determination of CD63 presence in sEVs obtained from the cell culture medium of young ADSCs. (D and E) Representative image of ADSC-sEVs as observed by transmission electron microscopy (TEM) and CD63 determination with immuno-gold labeling. (F) Images of kidney and muscle from old mice treated with PKH26-labeled sEVs from young ADSCs and vehicle as a control.

Fig 5: Exosomes secreted by metabolically impaired liver in T2DM migrate to periodontal tissue. (A) Cluster analysis of differentially expressed proteins (P value < 0.05 and Fold change >1.5) between db/m-exo and db/db-exo, with proteins abundance being Z-score normalized. (B) Volcano plots showed the fold change of different proteins. The green and red dots indicate down-regulated and up-regulated proteins, respectively. (C) The alveolar bone loss in db/m + Lig mice treated with exosome derived from different tissues was determined by micro‐CT. Scale bar, 200 μm. (D) Quantitative analysis of the CEJ-ABC in the micro‐CT (n = 6). (E) Histological evaluation of liver by H&E staining, Oil red O, Sirius red, and Fasn staining. Scale bars, 50 μm. (F) The expression of ASGPR on diabetic circulating exosomes analyzed by NanoFCM. (G) Immunoelectron microscopy detection of ASGPR and CYP2E1 in diabetic circulating exosomes. Scale bars, 100 nm. (H) The expression of ASGPR and Albumin was measured in circulating exosomes. (I) DiI-labelled liver-exosomes are internalized by CD90+ PDLCs in periodontal ligament. Scale bars, 20 μm. (J) The enrichment level of endogenous liver exosomes in periodontal bone site determined by in vivo imaging. H: heart; Li: liver; S: spleen; Lu: lung; K: kidney. (K) Fluorescence images showed the distribution of endogenous liver exosomes in periodontal ligament labelled by AAV-CD63-GFP. H: heart; Li: liver; S: spleen; Lu: lung; K: kidney. Scale bars, 50 μm. All results are representative data as mean ± SD. ns, not significant; **p < 0.01, ***p < 0.001 by one-way ANOVA. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)