Fig. 8

LUAD cells-derived exosomal LINC00963 induced M2 macrophage polarization.  Exosomes were isolated from the medium of A549 cells, observed for their morphology under TEM (A), measured for their size distribution by NTA (B), and analyzed for the expression levels of CD63, TSG101, CD9, GM130, calnexin by western blot (C). A549-derived exosomes were labeled with PKH67 and then incubated with macrophages. Macrophages were detected by immunofluorescence (D) or flow cytometry (E). Scale bar: 100 μm. F A549-derived exosomes 40 (µg/mL) were incubated with macrophages. Those incubated with IL-4 and IL-13 (20 ng/mL) were used as the positive control. Expression levels of CD206 and HLA-DR were examined by flow cytometry. G Exosomes isolated from indicated A549 cells were applied to macrophages. Expression levels of CD206 and HLA-DR on macrophages were examined by flow cytometry. H Exosomes were isolated from indicated A549 cells and examined for LINC00963 expression in exosomes by RT-qPCR. *P < 0.05, **P < 0.01, and ***P < 0.001