Journal: Food and Chemical Toxicology

Article Title: Nontoxic-dose deoxynivalenol aggravates lipopolysaccharides-induced inflammation and tight junction disorder in IPEC-J2 cells through activation of NF-κB and LC3B

doi: 10.1016/j.fct.2020.111712

Figure Lengend Snippet: Role of NF-κB in inflammation and intestinal epithelial barrier dysfunction of IPEC-J2 monolayers induced by LPS combined with nontoxic-dose DON. After corresponding treatment using DON and LPS, the expression of IκB-α, p p65 and p65 protein was determined by western blotting (A). The distribution of p65 inside and outside the nucleus was measured by immunofluorescence (B). Cells were immunostained with p65 (green) antibody and DAPI (nucleus, blue). After addition of inhibitor BAY 11–7082, the expression of TNF-α, IL-6 and TGF-β was measured by RT-qPCR (C) and the expression of NLRP3 (D) and tight junction protein (ZO-1 and Occludin) (E)was measured by western blotting. * P < 0.05 and ** P < 0.01 vs. control group. # P < 0.05 and ## P < 0.01 vs. LPS group. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Proteins were detected by specific primary antibodies: ZO-1 (AF5145, 1:1000, Affinity, China), Occludin (ab167161, 1:1000, Abcam, UK), NLRP3 (DF7438, 1:500, Affinity, China), procaspase-1 (AF5418, 1:500, Affinity, China), p65(AF1234, 1:1000, Beyotime, China), p p65 (AF5875, 1:1000, Beyotime, China), IκB-α (AF2176, 1:1000, Beyotime, China), LC3B (L8918, 1:1000, Sigma, USA).

Techniques: Expressing, Western Blot, Immunofluorescence, Quantitative RT-PCR, Control