Fig. 7.
Inhibitory effect of NCCoe-NGF-EVs on LPS-induced neuronal inflammation. A Confocal microscopy images of BV2 cells were acquired after treatment of DiR-labelled EVs from NCC and NCCoe-NGF lines for 6 h. Scale bar = 20 μm. B FACS analysis of BV2 cells treated DiR-labelled EVs from each cell lines for 6 h shows the EV uptake rate to BV2 cells. C Graph shows comparative expression level of Nitrite contained in the supernatant of LPS-induced BV2 cells after 6 h of pre-treatment with EVs derived from NCC and NCCoe-NGF lines. Data are shown as mean ± SD (n = 3). D ELISA assays were used to determine the protein levels of TNF-α (left panel) and IL-6 (right panel) in the cell supernatants of BV2 cells stimulated with LPS with or without each EV pre-treatment. Data are shown as mean ± SD (n = 3). E Western blot image shows the expression level of P-JNK, P-NF-kB, T-NF-kB, and β-actin in BV2 cells stimulated with LPS with or without each EV pre-treatment. F–G Fluorescence microscopy images confirming the expression levels of F-actin and Iba-1 in LPS-induced BV2 cells and a graph showing the relative immunoreactivity for Iba-1 expression in BV2 cells (relative to control). Data are shown as mean ± SD (n = 3). Scale bar = 20 μm. For multiple comparisons of groups, Statistical significance was determined using one-way ANOVA was performed followed by post hoc Tukey’s multiple comparison. *P < 0.05, **P < 0.01, ****P < 0.0001