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. 2023 Jul 9;13(12):3988–4003. doi: 10.7150/thno.84650

Figure 5.

Figure 5

DM-fBEVs are endocytosed by HK-2 cells and directly promote inflammation in vitro. (A) HK-2 cells were incubated with Dil-conjugated DM-fBEVs for 0, 2, 4, 12, 24 h. The uptake of fBEVs was observed by confocal laser scanning microscopy (scale bar, 2 μm, original magnification × 1000). (B-C) HK-2 cells were preincubated without or with inhibitors for 1 h and exposed to 5 μg/mL Dil-conjugated DM-fBEVs for 24 h. Cellular fBEVs were quantified (n=3, 5 pictures/group/experiment) (scale bar, 2 μm, original magnification × 1000). Control, cells without inhibitor treatment; Cyto, Cytochalasin D (1 μg/mL); Dan, Dansylcadaverine (200 μmol/L); Dyna, Dynasore (80 μmol/L); Nys, Nystatin (50 μmol/L). HK-2 cells were incubated with 5 μg/mL Control-fBEVs or DM-fBEVs for 24 h. (D-E) Protein (n=3) levels of IL-1β, TNF-α, MCP-1 and IL-6 were detected by Western blot analysis. (F) mRNA (n=4) levels of IL-1β, TNF-α, MCP-1 and IL-6 were measured by qRT-PCR analysis. *P < 0.05; **P < 0.01; ***P < 0.001.