FIGURE 5.

The biological effect of M2φ‐EVs was partly mediated by miRNAs. (a and b) The MH‐S cells treated with IL‐4 were transfected with NC siRNA or Dicer siRNA for 24 h. (a) Western blotting analysis of Dicer levels in MH‐S cells. The histogram showed the relative ratio of the expression levels of the target protein to that of an internal reference. (b) The mRNA of Dicer was detected by qPCR in MH‐S cells. (c–g) The MH‐S cells were stimulated with LPS (0.5 µg/mL) for 30 min and then treated with M2φ‐EVs, M2φ‐EVs^{NC siRNA} or M2φ‐EVs^{Dicer siRNA} (10 µg/mL) respectively for 23 h and then subjected to ATP (0.5 mM) for another 30 min. (c) Western blotting analysis of phosphorylation levels of p65 in MH‐S cells. The histogram showed the relative ratio of the expression levels of the target protein to that of an internal reference. (d) The concentrations of IL‐6, TNF‐α and IL‐1β were measured by ELISA in cell supernatant. (e) The mRNA expressions of IL‐6, TNF‐α and IL‐1β in MH‐S cells were analysed by qPCR. (f) Western blotting analysis of NLRP3, ASC and Pro‐caspase‐1 levels in MH‐S cells. The histogram showed the relative ratio of the expression levels of the target protein to that of an internal reference. (g) The mRNA expression of NLRP3 in MH‐S cells was analysed by qPCR. Representative results from three independent experiments are shown (n = 3). ns, not significant, *p < 0.05, **p < 0.01, ***p < 0.001 (one‐way ANOVA test excepted for unpaired Student t‐test in a, b; mean ± SD).