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Figure 5 | Journal of Neuroinflammation

Figure 5

From: Modulation of lipopolysaccharide-induced neuronal response by activation of the enteric nervous system

Figure 5

LPS-induced TNF-α production leads to TLR2 upregulation and TLR2 contributes to TNFα production. (A) Localization of TLR2 on neuronal or glial cells of the rENSpc was analyzed by immunocytochemistry using anti-TLR2, –Tuj (β-tubulin III) and –S100β antibodies on cultures treated for seven hours with LPS (0.1 μg/ml), or not. Scale bar: 50 μm. (B) Expression of TLR2 was also observed in human EGC cultures by immunocytochemistry using anti-TLR2 and –S100β antibodies on cultures treated for seven hours with LPS (0.1 μg/ml), or not. (C) Relative expression of TLR2 mRNA in rENSpc treated (+), or not (−), with LPS for seven hours (15 independent samples). Values represent the mean ± SEM of TLR2 mRNA quantification against RPS6 (Mann-Whitney U test; *P <0.05 as compared with control without LPS). (D) Quantification of TLR2 mRNA in rENSpc treated (+), or not (−), with LPS (0.1 μg/ml) in the presence of, or not, anti-TNF-α (1 μg/ml and 10 μg/ml), was performed by RT-qPCR (five independent samples) (Mann-Whitney U test; *P <0.05 as compared with control without LPS; #P <0.05 as compared with LPS treatment without anti-TNFα). (E) TLR2 mRNA expression in rENSpc treated (+), or not (−), with LPS in the presence of, or not, EFS stimulation (11 to 16 independent samples). TLR2 mRNA quantification relative to S6 was expressed as induction fold to the control mean (Mann-Whitney U test; *P <0.05 as compared with control without LPS; #P <0.05 as compared with LPS treatment without EFS). (F) ENS cultures were treated (+), or not (−), with LPS (0.1 μg/ml) and Pam3CSK4 (TLR1/2 agonist; 0.1 μg/ml). Values represent the mean ± SEM of eight independent experiments (Mann-Whitney U test; *P <0.05 as compared with control without LPS; §P <0.05 as compared with Pam3CSK4 without LPS).

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