Fig 1: Levels of VTN in the tears and conjunctiva are elevated in DED mice. (a) Tear fluid and serum concentrations of VTN in DED mice were assessed using ELISA. Data (mean ± SE) are obtained from 10 mice from day 0 through 3 weeks. * p < 0.001 versus day 0. (b) Immunohistochemical staining of VTN (brown color) in the conjunctiva. Representative micrographs from two independent experiments (n = 6) are shown. Analysis of VTN expression in the conjunctiva of DED and NS mice (each n = 6) by qPCR (c) and Western blotting (d). Representative blots and densitometric analyses of 6 samples are shown. a p < 0.0001 versus NS mice. b p < 0.0002 versus NS mice.
Fig 2: VTN is a macrophage proinflammatory stimulator. (a) qPCR analysis of the proinflammatory cytokine expression induced by NaCl for 24 h. Data were normalized to gapdh expression levels and summarized as the mean ± SE of three separate experiments. (b–d) VTN promotes the expression of the TNF-a, IL-1ß, andIL-6genesin NaCl-treated BMDMs. Cells were cultured in isomolar medium or hyperosmotic medium by adding 20 mM NaCl for 24 h and were then treated with VTN for a further 3 h before qPCR analysis. Data are from three independent experiments. a p < 0.05 versus VTN untreated cells. b p < 0.01 versus corresponding UT/VTN-treated cells.
Fig 3: VTN induces NF-?B activation in NaCl-treated BMDMs to upregulate proinflammatory cytokine gene expression. BMDMs were treated with 20 mM NaCl for 24 h and were then treated with 20 µM c(RGDfK), 10 µM SN50, 5 µg/mL of anti-Itg av antibody, or IgG control for 10 min prior to stimulation with 200 ng/mL of VTN for a further 3 h. (a) Western blotting of NF-?B p65 phosphorylation (p-p65) and total p65 stimulated by VTN for 20~60 min. Representative blots and densitometric analyses from three samples are shown. a p < 0.001versusuntreated cells. b p < 0.005 versus corresponding VTN-treated cells. (b) qPCR analysis of proinflammatory gene expression stimulated by VTN for 3 h. Data were summarized as mean ± SE of three separated experiments. (c) Representative Western blots of p-p65 and total p65 stimulated by VTN for 20 min and densitometric analyses from three samples are shown. c p < 0.002 versus Cont Ab-treated cells. (d) qPCR analysis of proinflammatory gene expression stimulated by VTN for 3 h. Data were summarized as the mean ± SE of three separated experiments. d p < 0.0001 versus Cont Ab-treated cells.
Supplier Page from Wuhan Fine Biotech Co., Ltd. for Mouse Vtn(Vitronectin) ELISA Kit