Fig 1: Blocking the Notch signaling inhibits the activation of the NF-κB pathway. (A): p65 levels in mouse hearts were detected by immunohistochemistry staining; (B): Phosphorylation level of the NF-κB pathway was detected by Western blot; (C): mRNA expression of CYLD in mouse cardiac macrophages was detected by qRT-PCR; (D): Protein expression of CYLD in mouse cardiac macrophages was detected by Western blot; N = 6; three independently repeated tests were performed, and the data were expressed as mean ± SD. One-way ANOVA was used for the analysis of variance, and Tukey's multiple comparisons test was used for post hoc test; ** P < 0.01 (compared with the sham group), and ## P < 0.01 (compared with the sh-NC group).
Fig 2: RSV and RELA regulate intracellular N-glycosylation of the integrin signaling pathway. N-glycoproteins were identified in cytoplasmic proteins from control and RSV-infected cells in the presence or absence of RELA silencing. (A) Agglomerative hierarchical clustering of 171 peptides significantly changed across the treatment conditions. Data was Z-scored across the row prior to clustering using Euclidian distance. The legend is at right. Each row represents an individual peptide; columns refer to different cell treatments/conditions. Note that the replicates for each treatment group cluster together and that the bulk of the N-glycopeptides are induced by RSV infection and are not observed in the RELA-depleted cells. (B) Volcano plot of peptides in RSV- versus mock-treated samples. X axis, fold change RSV versus mock; Y axis, log2 (ANOVA). Selected peptides in the integrin signaling pathway are labeled. (C) Volcano plot of peptides in RSV-infected cells WT versus RELA knockdown samples. X axis, fold change RSV versus mock; Y axis, log2 (ANOVA). Selected peptides in the integrin signaling pathway are labeled. (D) LFQ intensity for selected integrin (ITG), galectin-3 binding protein (LGALS3BP), ICAM1 and laminin subunit gamma 1 (LAMC1) peptides. **, p < 0.01 post-hoc.
Fig 3: OGG1 inducibly binds to HBP regulatory elements and recruits phospho-Pol II. XChIP was performed on control or RSV-infected hSAECs; data are expressed as a fold change over IgG control. (A) OGG1 binding to the GFPT2 intragenic enhancer. (B) OGG1 binding to the UAP1 promoter. (C–F), Cells were RSV infected in the presence of solvent (DMSO) or TH5487 as indicated. (C,D), XChIP for RELA binding. (E,F), XChIP for phospho-Ser2 CTD Pol II binding. *, p < 0.05; **, p < 0.01; n.s., not significant. Each symbol is an independent immunoprecipitation.
Fig 4: Effects of lncRNA CASC2 on caspase-cascades and the NF-κB pathway. (A) A schematic diagram showing two different modes of TRAIL acting on TRAIL-sensitive and TRAIL-resistant cell lines. (B) Huh-7 and HCCLM3 cells were exposed to 1, 10, 100, and 1,000 ng/ml rhTRAIL protein and examined for the cell viability by MTT assay. The IC50 values were calculated and shown. Regular Huh-7 and HCCLM3 cells were then divided into TRAIL-sensitive Huh-7 (S) and HCCLM3 (S) and TRAIL-resistant Huh-7 (R) and HCCLM3 (R). (C) Huh-7 (S), HCCLM3 (S), Huh-7 (R), and HCCLM3 (R) cells were treated with 0 or 150 ng/ml rhTRAIL and examined for the protein levels of RIPK1, caspase-8, caspase-3, IKKβ, p-IκBα, and p-p65 using Immunoblotting. (D) CASC2 expression was determined in Huh-7 (S), HCCLM3 (S), Huh-7 (R), and HCCLM3 (R) cells using qRT-PCR. (E) Huh-7 (S) and HCCLM3 (S) cells were transfected with CASC2 or sh-CASC2 and examined for the protein levels of caspase-8 and caspase-3 using Immunoblotting. (F) Huh-7 (R) and HCCLM3 (R) cells were transfected with CASC2 or sh-CASC2 and examined for the protein levels of IKKβ, p-IκBα, and p-p65 using Immunoblotting. **p < 0.01, compared with sensitive group or vector-NC group, ##p < 0.01, compared with sh-NC group.
Fig 5: Melatonin (Mel) alleviated the degeneration of endplate cartilaginous cells (EPCs) through inhibiting inflammatory response and matrix degradation activated by NF-κB pathway. (A–C) qRT-PCR analysis of the relative mRNA expression of IL-1β (A), matrix metalloproteinase-3 (MMP3) (B), and ADAMTS-5 (C) under lipopolysaccharide (LPS), Mel, and Luzindole treatment. (D) Western blotting for proteins of IL-1β, MMP3, ADAMTS-5, p-P65, and P65 under LPS, Mel, and Luzindole treatment. (E–H) Quantitation of relative protein level of IL-1β, MMP3, ADAMTS-5, and p-P65. (I) Immunofluorescence staining of P65 nuclear translocation under LPS, Mel, and Luzindole treatment (∗P < 0.05, ∗∗P < 0.01).
Supplier Page from Abcam for Anti-NF-kB p65 (phospho S276) antibody