Figure 5.

Knockdown of lincRNA-Cox2 Restores EV-Mediated Impairment of Microglial Phagocytosis

(A) Real-time PCR for lincRNA-Cox2 in microglia transfected with lincRNA-Cox2 siRNA. (B) Heatmap of differentially expressed phagocytosis-associated genes in lincRNA-Cox2 knockdown BV-2 cells compared with control cells. (C) Mouse primary microglial cells were transfected with lincRNA-Cox2 siRNA for 24 hr, followed by exposure of cells to astrocyte EVs for 4 hr, and subsequently assessed for the expression of Lrp, Pld2, and Syk by qPCR. (D and E) Mouse primary microglial cells were pre-treated with TLR7 inhibitor (chloroquine 25 μM) or NF-κB inhibitor (SC-514, 5 μM) for 1 hr, followed by exposure of cells to astrocyte EVs and subsequent exposure to fluorescent beads for 2 hr. Phagocytosis was measured using (D) fluorescence microscopy (scale bar, 20 μm; high-magnification images are shown in the bottom right corners) and (E) fluorometric plate reader. (F) TLR7 KO microglial cells were exposed to astrocyte-derived EVs and assessed for phagocytosis as described above. All data are presented as mean ± SD. *p < 0.05 versus control; ^#p < 0.05 versus treatment using Student’s t test.