Fig 2: The response of NFκβ members to increased DLG2 expression and activation of the inflammasome in THP-1 cells. The expression of a RELA, b RELB, c NFKB1 and d NFKBIZ in response to control conditions (circle symbol), LPS priming (square symbol) and LPS + ATP treatment (triangle symbol) with or without DLG2 overexpression. e Representative immunoblot showing the expression of; DLG2, p-P65 (Ser 536), RELA, RELB, p-P105 (Ser 932), NFκB1, IκBζ and GAPDH. Each experiment was performed in triplicate. The gene expression data are presented as log2 fold change and plotted as mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001

Fig 3: Interaction between ANXA4 and NF-?B p50 in OCCC cell lines. (A) Co-immunoprecipitation analysis was performed to determine the association between ANXA4 and NF-?B p50. (B) Double-labeling immunofluorescence analysis demonstrated co-localization of ANXA4 and NF-?B p50 in OCCC cells. (C) Western blot analysis was performed to detect ANXA4 and NF-?B p50 expression following transfection of the ANXA4 gene. (D) Immunocytochemistry analysis was performed to detect ANXA4 and NF-?B p50 expression following transfection of the ANXA4 gene. *P<0.05. ANXA4, Annexin IV; NF-?B, nuclear factor-?-light-chain-enhancer of activated B cells; OCCC, ovarian clear cell carcinoma; IB, immunoblotting; IP, immunoprecipitation; NOT, anti-IgG antibody (negative control).

Fig 4: Expression levels of ANXA4, NF-κB p50 and mutant p53 in ovarian clear cell carcinoma tissues (magnification ×400). ANXA4, Annexin IV; NF-κB, nuclear factor-κ-light-chain-enhancer of activated B cells.

Fig 5: IL-1/OSM activate RelB/p50 in GBM.a Primary GBM12 cells and established U373 and U87 cell lines were stimulated with IL-1/OSM as indicated. Expression of RelB, p65, p105, p50, p100, and p52 was analyzed by western blotting. Tubulin was used as a loading control. b U373 cells were treated with IL-1/OSM as indicated, expression was assessed by western blotting. c Nuclear and cytosolic fractions were prepared from control or IL-1/OSM-stimulated cells (at indicated time points), and RelB expression was analyzed by western blotting. Lamin A/C and tubulin were used to examine purity of the fractions. d Cells were stimulated for 18 h, and RelB visualized by immunofluorescence. DAPI was used to stain nuclei. e Cells transfected with the indicated siRNAs were stimulated 48 h later with IL-1/OSM for 18 h. Expression was analyzed by qPCR (three experiments, error bars represent s.d., *p < 0.05 (two-way ANOVA, Sidak’s test). f U373 were stimulated for 8 h, RelB was immunoprecipitated, and p50 and RelB were detected by western blotting. g Binding of RelB, p50, and p52 at the indicated cytokine promoters was analyzed by ChIP. U373 cells were stimulated with IL-1/OSM for 8 h. Normalized binding is shown. IgG was used as a control for IP (dotted line). n = 3–6, error bars represent s.d., *p < 0.05 (T-test, Sidak’s test). h Expression was analyzed by qPCR in human GBM tumors (n = 6) and normal brains (n = 4). i RelB was visualized in tumor sample of GBM patient by immunofluorescence. DAPI was used to counterstain nuclei. j Genomic copy number analysis of NF-?B family members, GISTIC data downloaded from TCGA via cBioPortal (n = 206)