Fig. 1

LEPCs suppress LPS-induced exosome release from Mϕ. a Exosomes were isolated from BMDM cultured in serum-free medium, stained with PE-isotype and PE-CD63 antibodies and measured by flow cytometry. b Exosomes isolated from the culture media of control and LPS-treated BMDM were analyzed for the mean particle diameter and total number by NanoSight. c BMDM were treated with LPS (1 μg/ml) for 0, 6, 12, and 24 h. Exosomes were then isolated from the culture medium and detected by CD63 staining and flow cytometry. d BMDM were treated with LPS (1 μg/ml) in the presence or absence of dimethyl amiloride (DMA, 25 μM) for 24 h. Exosomes were then isolated from the culture medium and detected by CD63 staining and flow cytometry. e Lung cells were labeled with biotin-conjugated CD326 antibody and separated with streptavidin-conjugated immunomagnetic beads. Flow cytometric analysis shows a purity of greater than 95% for the yielded LEPCs. f BMDM were cultured alone or cocultured with LEPCs and were either treated with LPS (1 μg/ml) for 24 h or not treated. Exosomes were isolated from the culture medium and detected by CD63 staining and flow cytometry. All results are representative of at least three independent experiments. The graphs show the mean ± SEM, n = 3; *P < 0.05 or **P < 0.01, compared with the indicated groups