Fig. 5
Exosomal miR-155-5p promotes macrophage M1 polarization and inflammation via activating p38MAPK in vitro. A Uptake of PKH67-labeled exosomes (green) by RAW264.7 macrophages co-incubated for 6 h, observed under inverted fluorescence microscopy. B Expression of miR-155-5p in RAW264.7 macrophages co-incubated with iHvKp-exo or control exosomes for 48 h, measured by RT-PCR. C Levels of pro-inflammatory cytokines in co-incubated cells after 48 h, measured by RT-PCR. D Level of M1 polarization measured by flow cytometry. E The relationship between exosome uptake and M1 polarization of macrophages was assessed using flow cytometry. Exosomes were labeled with PKH26, while M1 macrophages were labeled with FITC-CD86 antibody. F Activation of p38MAPK by exosomes measured by Western blotting. P-p38MAPK/Total p38MAPK is used to assess the phosphorylation level of p38MAPK. G Activation of p38MAPK in RAW264.7 macrophages transfected with miR-155-5p inhibitor or NC inhibitor before co-incubation with iHvKp-exo, evaluated by Western blotting. P-p38MAPK/Total p38MAPK is used to assess the phosphorylation level of p38MAPK H Pro-inflammatory cytokines expression levels measured by RT-PCR. I Level of M1 polarization measured by flow cytometry. Data are represented as mean ± SEM, *p < 0.05, **p < 0.01, ***p < 0.001, n = 3