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Fig. 6 | Journal of Neuroinflammation

Fig. 6

From: Anti-inflammatory role of GM1 and other gangliosides on microglia

Fig. 6

Inhibition of the ganglioside biosynthetic pathway enhances microglial response to LPS. A Dot-blot and relative quantification of cellular ganglioside levels in BV2 cells treated with the indicated concentrations of GENZ-123346 for 72 h shows a significant decrease in levels of GM1 and GD1a (N = 3). B Representative histogram and relative flow cytometry quantification (mean fluorescence intensity, MFI) of TLR-4 present at the plasma membrane of BV-2 cells after treatment with 10 µM GENZ-123346 for 72 h. C Representative immunoblot showing increased phosphorylation of IKK and p38 MAPK in BV-2 cells treated with GENZ-123346 and stimulated with LPS (100 ng/ml) for the indicated time. The numbers under the blots indicate the fold-change of p-IKK and p-p38-MAPK compared to untreated control, after normalization over total IKK or p38-MAPK protein levels. D Expression of IL-1β and TNF mRNA in BV-2 cell stimulated with LPS (100 ng/ml) after 72 h incubation with GENZ-123346 (5 μM) (N = 3). E Dot-blot and quantification of ganglioside levels in primary mouse microglia incubated with 10 µM GENZ-123346 for 72 h show a significant decrease in the levels of both GM1 and GD1a (N = 3). F Expression of IL-1β and TNF mRNA in mouse microglia treated with GENZ-123346 as in E, and after stimulation with the indicated concentrations of LPS for 6 h (N = 3). G TNF secretion by control and GENZ-123346-treated cells upon stimulation with LPS (1 ng/ml) (N = 3). Data shown are mean values ± STDEV. One-way ANOVA with Sidak’s multiple comparisons test was used in A; two-tailed t test was used in B and E; two-way ANOVA with Tukey’s multiple comparisons test was used in D, F and G. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001

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