Fig. 2: β cells secreted small EVs containing miR-503-322 to enter acinar cells. | Nature Communications

Fig. 2: β cells secreted small EVs containing miR-503-322 to enter acinar cells.

From: Endocrine-exocrine miR-503-322 drives aging-associated pancreatitis via targeting MKNK1 in acinar cells

Fig. 2

A–C Identification of EVs collected from 20-week-old WT and βTG mouse islet β-cells: TEM images showing the morphology of βEVs collected (A), NTA analysis of nanoparticle size distribution (B), and molecular marker identification of βEVs (C). n = 2 independent experiments. As there was no significant difference between WT and βTG, the results are presented in the figure for βEVs from WT β-cells. D NTA analysis of nanoparticle concentration of βEVs collected from 20-week-old WT and βTG mouse islet β-cells. n = 3. E qPCR analysis of miR-503 expression in βEVs of WT and TG. n = 3. F PKH67-labeled WT-βEVs or βTG-βEVs were co-incubated with fresh acini for 8 h. Representative confocal images of PKH67 (green), phalloidin (red), and nuclei (blue) in primary acini and quantification of PKH67 fluorescence intensity. n = 3 replicates, with each data point representing a count of 3 acini. G qPCR analysis of miR-503 expression in the acini of received βEVs. n = 3 independent experiments. H, I Experimental scheme: PKH67-labeled WT-EVs or βTG-EVs were infusion into the C57BL/6 J male mouse pancreas via pancreatic ductal. The pancreas was harvested after 12 h, stained with frozen sections for amylase, and then visualized. Representative confocal images of PKH67 (green), amylase (red) and nuclei (blue) of pancreas and quantitation of relative PKH67 fluorescence intensity. n = 3 mice. J–L Pancreatitis parameters assay after initial caerulein (50 μg/kg) injection 7 h in 12-week-old WT and β-cell-specific miR-503-322 knock-in (βKI) female mice. H&E and F4/80 immunohistochemistry (IHC) of pancreatic sections, quantitation of the number of F4/80 positive cells in pancreatic sections under ×200 microscopic view (J), pancreatic histological scores (K) and level of serum amylase (L). Arrows indicate the macrophages. n = 5 mice. Data are means ± SEM. Unpaired Student’s t tests were used to evaluate statistical significance. Source data are provided as a Source Data file.

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