Figure 7.

Vav1 knockdown represses inflammation in BV-2 cells subjected to OGD/R.

(A) IL-1β, TNF-α, and IL-18 mRNA levels in control BV-2 cells and BV-2 cells subjected to OGD/R were analyzed by qPCR. (B) The contents of IL-1β, TNF-α, and IL-18 in control BV-2 cells and BV-2 cells subjected to OGD/R were detected by ELISA. (C) Immunofluorescence staining for NLRP3 (FITC, green) in BV-2 cells subjected to OGD/R. Compared with control cells, NLRP3 expression was increased in BV-2 cells subjected to OGD/R, while inhibition of Vav1 expression decreased NLRP3 expression. Nuclei were stained with DAPI (blue). Scale bars: 50 μm. (D) Quantification of NLRP3 positive cells. (E, F) NLRP3, ASC, and cleaved caspase-1 (p20) protein levels in BV-2 cells subjected to OGD/R were analyzed by western blot. Data are expressed as the mean ± SD, and were analyzed by one-way analysis of variance followed by Tukey’s multiple comparisons test. The experiments were repeated three times. ASC: Apoptosis-associated speck-like protein; DAPI: 4′,6-diamidino-2-phenylindole; ELISA: enzyme-linked immunosorbent assay; FITC: fluorescein isothiocyanate; IL-18: interleukin-18; IL-1β: interleukin-1β; NLRP3: NOD-like receptor pyrin 3; OGD/R: oxygen-glucose deprivation/reoxygenation; qPCR: quantitative polymerase chain reaction; TNF-α: tumor necrosis factor-α; Vav1: Vav guanine nucleotide exchange factor 1.