Fig. 2

dNPc-exo induces M1 polarization of macrophages via miR-27a-3p. (A) Representative images of immunofluorescence staining of F4/80 and iNOS in M0 macrophages after dNPc-exo and nNPc-exo incubation. Scale bar = 100 μm. (B) Quantitative analysis of the percentage of iNOS⁺ macrophages in immunofluorescence staining. (C) RT-qPCR analysis of iNOS gene expression in macrophages with nNPc-exo and dNPc-exo treatment. (D) Western blot analysis of iNOS expression in macrophages with nNPc-exo and dNPc-exo treatment. (E) Quantitative analysis of iNOS expression in western blot. (F) Flow cytometry analysis of the macrophages. (G) Quantitative analysis of the iNOS⁺ macrophages percentage in flow cytometry. (H) Heatmap of nNPc-exo group and dNPc-exo group. (I) Cluster analysis of RNA-Seq between nNPc-exo group and dNPc-exo group. (J) A volcanic map of the differentially expressed genes between nNPc-exo group and dNPc-exo group. (K) Pathway enrichment factor map of differential genes (Top20). (L) RT-qPCR analysis of seven miRNAs that highly expressed in dNPc-exo group. (M) RT-qPCR analysis of iNOS gene expression in M0 macrophages with different miRNAs mimic treatment. The data are expressed as the mean ± SEM. n = 3. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001; ns, non-significant difference