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Fig. 4 | Microbiome

Fig. 4

From: Non-digestible oligosaccharides directly regulate host kinome to modulate host inflammatory responses without alterations in the gut microbiota

Fig. 4

Functional validation of NF-κB pathway in IECs. a The NF-κB network with the top five predicted nodes generated from GeneMania. b Caco-2Bbe1 cells were treated with inulin or scFOS (10% w/v, 16 h) before 3 h challenge with EHEC (MOI of 10:1). Cells were then lysed and immunoblotted to check for NF-κB-p65 phosphorylation (n = 4) and IKBα degradation (n = 5). c Localization of NF-κB-p65 in prebiotic-treated Caco-2Bbe1 cells upon 3 h EHEC challenge or 15 min TNF-α challenge (arrows indicate nuclear translocation, representative of 3 separate experiments). d, e Inulin and scFOS decreased EHEC and TNF-α-triggered IL-8 expression (n = 3). f, g Caco-2Bbe1 cells pre-exposed to inulin or scFOS (10% w/v) were challenged with TNF-α and IFN-γ (20 ng ml−1 each, 30 min), TNFα alone (20 ng ml−1, 15 min) and LPS (200 ng/ml). Cells were lysed and immunoblotted for IKBα levels and GAPDH loading control (n = 4). Western blot bands were cropped from the original blots of each individual experiment. Bars represent means ± SEM, *P < 0.05 (ANOVA Bonferonni post hoc test)

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