Fig. 2

NAC-derived tumor exosomes increase miR-155-5p level in macrophages. A. THP-1 cells were treated with PMA at 100 nM for 24 h. The macrophage markers CD163 and CD206 were assessed by immunoblotting. B. Exosomes isolated from A2780 cells was labeled using lipophilic tracer DiR before being incubated with macrophages for 24 h. Cellular uptake of exosomes were examined by confocal microscopy. Macrophage nuclei were labeled with DAPI and actin cytoskeleton was labeled with Phalloidin. C. MΦ was treated with A2780-derived exosomes (Exo-con) and NAC-treated A2780-derived exosomes (Exo-NAC) at 100 μg/ml for 48 h. MiR-155-5p level was detected by RT-qPCR. D-G. Macrophages were transfected with DICER siRNASmartpool (si Dicer) or negative siRNA control (si Cont) for 24 h followed by Exo-con and Exo-NAC treatment for another 48 h. (D) Dicer mRNA expressions in macrophages were determined. (E) miR-145 and miR-9 expression levels were set as controls. (F, G) Pre-miR-155 and miR-155-5p levels were measured by qPCR in macrophages with treatments as indicated. Data are representative of three independent experiments. Data = Mean ± SD. *p < 0.05, **p < 0.01