Fig. 3

ASH-WEX and FIV pretreatment inhibits LPS-induced various inflammatory pathway protein expression in BV-2 microglia. A Representative confocal images of activated BV-2 microglial cells with or without pretreatment with ASH-WEX and FIV immunostained with (a) Iba-1, (b) NFkB, and (c) AP1. B i Representative Western blots of NFkB and AP1 showing inhibition of the translocation of the NFkB from cytoplasm to nucleus and decrease in expression of AP1 with ASH-WEX and FIV pretreatment. B, iii, iv Representative histograms present normalized relative expression of NFkB and AP1 plotted as mean ± SEM calculated from three independent experiments. Cytoplasmic expression of protein was normalized with α-tubulin, and nuclear expression was normalized with histone-3. B ii Representative Western blots of pAkt, OX-6 showing decrease in expression of pAkt, OX-6 with ASH-WEX and FIV pretreatment. B v Histogram representing mean of densitometric analysis of Western blot signals of pAkt, OX-6 after normalization with α-tubulin. C ASH-WEX and FIV inhibit the release of the inflammatory mediators due to neuroinflammation induced by LPS treatment. C i Represents the relative production of the nitrite in different treated groups as compared to control due to neuroinflammation as estimated by the Griess reagent. C ii Represents histograms depicting the decreased levels of the inflammatory cytokines (TNF-α, IL-1β, IL 6) by the ASH-WEX and FIV treatment as estimated by Sandwich ELISA. C iii Representative MMP zymograms for control and different treated groups and histograms represent densitometric analysis of MMP bands. *p < 0.05 represents statistically significant difference between control and LPS + ASH-WEX and LPS + FIV-treated groups. ⁺ p < 0.05 represents statistically significant difference between control and LPS-treated group. ^# p < 0.05 represents statistically significant difference between LPS and LPS + ASH-WEX and LPS + FIV-treated groups