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. Author manuscript; available in PMC: 2023 Dec 14.
Published in final edited form as: Gut. 2022 Jan 11;71(11):2233–2252. doi: 10.1136/gutjnl-2021-326269

Figure 2.

Figure 2

Germ-free (GF) 3xTg mice demonstrate attenuated C/EBPβ/AEP pathway and AA-associated inflammation. (A) immunoblot showing p-C/EBPβ, C/EBPβ, AEP, APP and tau expression and processing, as well as arachidonic acid metabolism in mouse brains of germ-free 3xTg mice and SPF 3xTg mice. (B) Quantitative analysis of immunoblot. The bands of active-AEP, TauN368, APPN585, C/EBPβ, LOX5, Cox1, COX2, BLT1 and BLT2 were measured with image J and normalised with β-actin. n=5 in each group, data are shown as mean±SEM, *p<0.05, **p<0.01 compared with control, unpaired t-tests. (C) AEP activity assay in the brain lysates from germ-free 3xTg mice and SPF 3xTg mice. SPF 3xTg mice showed escalation in AEP activities compared with germ-free 3xTg mice. Data represent the mean±SEM; representative data of five samples; **p<0.01 compared with control, unpaired t-tests. (D) proinflammatory cytokine IL1-β, IL-6 and TNFα concentrations in the brain lysates from germ-free 3xTg mice and SPF 3xTg mice, respectively. Data represent the mean±SEM; representative data of five samples; ***p<0.001 compared with control, multiple unpaired t-tests. (E) Quantitative RT-PCR analysis of the brain samples from GF mice versus spf mice comparing C/EBPb targeted AA pathway genes. AA, arachidonic acid; AEP, asparagine endopeptidase; COX, cyclooxygenases; SPF, specific-pathogen-free.