Fig. 3

ADSC-EVs decrease LPS-stimulated activation, TLR4/MAPK/NF-κB proinflammatory cascade signaling in BV2 cells induced by LPS. (A) Representative images of the uptake of Dil-labeled EVs (red) by BV2 cells. (B) Representative immunostaining images and quantification of CD68 in LPS-activated BV2 cells after culturing with PBS or ADSC-EVs for 24 h (n = 6, ***p < 0.001). (C) Representative flow cytometry images and iNOS fluorescence intensity quantification in LPS-activated BV2 cells after culturing with PBS or ADSC-EVs for 24 h (n = 3, **p < 0.01, ***p < 0.001). (D) Quantification of IL-1β, IL-6, and TNF-α mRNA levels relative to 18s RNA by qRT-PCR in LPS-activated BV2 cells after culture with PBS or ADSC-EVs for 24 h (n = 3, *p < 0.05, **p < 0.01, ***p < 0.001). (E) Quantification of IL-1β, IL-6, and TNF-α production in LPS-stimulated BV2 cell culture medium after treatment with ADSC-EVs or PBS for 24 h (n = 4, *p < 0.05, ***p < 0.001). (F) Western blot images of IL-6, TNF-α, and IL-1β relative protein expression normalized to tubulin levels (n = 3, *p < 0.05, **p < 0.01, ***p < 0.001). (G) Western blot images of TLR4, p-NFκB, p65, p-p38 MAPK, and p-ERK relative protein expression normalized to Western blot images of IL-6, TNF-α, and IL-1β relative protein expression normalized to tubulin levels (n = 3, *p < 0.05, **p < 0.01, ***p < 0.001)