Fig 2: Nuclear factor (NF)-?B partially mediates neutrophil activation induced by post-I/R lymph. (A) Post-I/R lymph, but not pre-I/R lymph, activates NF-?B. n = 6/group. (B) Post-I/R lymph induces NF-?B nuclear translocation. Red arrows indicate nuclear phospho (p)-p65. Representative images are from 3 independent experiments. Scale bar, 20 µm. (C) Blocking NF-?B activity attenuates neutrophil Tnf and Cxcl5 gene expression induced by post-I/R lymph. NF-?B i, NF-?B inhibitor (1 µM). n = 6/group. *p < 0.05, **p < 0.01 vs. control and ##p < 0.01 vs. post-I/R lymph. (D) Gut I/R induces neutrophil NF-?B activation, which is inhibited by MLDL. n = 5–6/group. (E) Gut I/R upregulates the expression of Tnf and Cxcl5 by lung neutrophils, which is suppressed by MLDL. *p < 0.05, **p < 0.01 vs. sham or MLDL + sham and #p < 0.05 vs. I/R. One-way ANOVA was used.

Fig 3: Cotreatment with EGCG and BAY11-7082 significantly inhibits NF-?B signaling. (A-D) Western blot analysis of total NF-?B and phosphorylated NF-?B p65 in A549 and H1299 whole cell lysates after treatment with 20 µM EGCG; 0.625, 1.25, or 2.5 µM BAY 11-7082; or the combination thereof for 48 h. (E) A549 and H1299 cells were cotransfected with SV40 and SV40 for dual luciferase reporter assays. (F) qRT-PCR analysis of NF-?B expression levels. (G-H) qRT-PCR analysis of C-MYC, Cyclin D1, Bcl-2, Bcl-xL, COX-2, TNF-a, TWIST1, and MMP2 mRNA expression in A549 and H1299 cells after treatment with 20 µM EGCG; 0.625, 1.25, or 2.5 µM BAY11-7082; or the combination thereof for 48 h. Data are shown as the mean ± S.D. (n=3). * P < 0.05, ** P < 0.01, ***P< 0.001.

Fig 4: circCCDC134 stimulates HIF1A transcription by recruiting p65. A The molecular simulation results supported that circCCDC134 could perfectly dock p65. The results of B: circRNA pull-down and C: RIP assays demonstrated that p65 could interact with circCCDC134, and D: no significant change was found in the p65 protein levels upon circCCDC134 knockdown. E Heatmaps and G: pie charts of p65 ChIP-seq. HIF1A was found to be the key target gene of p65. H and I p65 was enriched in the region - 1200 to - 400 bp from the transcription start site of the HIF1A promoter. p65 enrichment of the HIF1A promoter was reduced after interfering with J: p65 and K: circCCDC134 expression. L The expression of HIF1A was positively correlated with the expression of circCCDC134. M and N Immunohistochemistry showed that HIF1A was positively correlated with the expression of circCCDC134 in the xenograft growth and lung metastasis models

Fig 5: CBD treatment inhibited NF-?B pathway activation induced by the EHFD diet. Mice were fed ethanol plus high-fat high-cholesterol diet for 8 weeks with or without CBD treatment. Control mice were fed a standard chow diet. Western blot analyses of pI?Ba, I?Ba, pNF-?B p65, and NF-?B p65 (A). Quantification of expressions of I?Ba, pI?Ba, and pNF-?B p65 (B). Values represent mean ± SEM (n = 6–8/group). *p < 0.05 vs the control group; #p < 0.05 vs the EHFD group.