Fig. 3

The intracellular uptake and anti-inflammatory effects of MCEVs. (A) Fluorescent staining to show cellular uptake of MCEVs after 4, 8, and 12 h. MCEVs were labeled with red DiL dye. The cytoskeleton was stained with green FITC-Phalloidin dye. The nucleus was stained with blue DAPI dye. (B) A representative image of the cellular uptake efficiency of MCEVs shown by flow cytometry. (C-E) qRT-PCR to evaluate the transcription level of inflammatory cytokines by RAW 264.7 cells after incubation with MCEVs for 12 h and then 500 ng/mL LPS induced for 12 h. (F-H) RAW 264.7 cells were incubated with 10 µg/mL MCEVs for 12 h and then induced by 500 ng/mL LPS for 12 h. (F) Representative images of CD86 antibody immunofluorescence staining of M1 macrophages. (G) Flow cytometry was used to detect the level of ROS by DCFH-DA ROS probe. (H) Statistical map of DCF-positive cells. Data were shown as mean ± SD. n ≥ 3, *: P < 0.05, **: P < 0.01, ***: P < 0.001, ns: P > 0.05, indicating no significance