Fig 2: Diapocarotene-dials attenuate the NF-?B signal in RANKL-activated RAW264.7 cells. 5 × 106 cells in 100-mm plates were either incubated alone or in the presence of the indicated diapocarotene-dials (10 µM) or lycopene (10 µM) for 2 h, and then treated with RANKL (40 ng/mL) for 15 min (a,b) and 40 min (c–g). Whole cell lysates (a,b) and cytoplasmatic and nuclear fractions (c–g) were prepared and analyzed by Western blotting, as described in Materials and Methods. Values are the means ± SE of three independent experiments. (a) Blots of I?Ba and pI?B. (b) The ratio of p-I?B:I?B after normalization with calreticulin (cal) is presented as the % of the control without RANKL. *** p < 0.001 for the difference between RANKL to the control. * p < 0.05 for the difference between RANKL to 6,14' and 10,10'. (c) Blots of nuclear and cytosolic p65. (d) Nuclear p65 levels normalized to laminin B. (e) Blots of nuclear and cytosolic p52 and p100. (f) Nuclear p52 levels normalized to laminin B. Results are % of RANKL. (g) Nuclear p100 levels normalized to laminin B. Results are % of control.

Fig 3: Representative IHC staining for p52 and p65 expression, the VUMC TROC. Representative immunohistochemical (IHC) staining for p52 and p65 in a variety of ovarian tumor histologic subtypes. Staining was predominantly cytoplasmic, with a smaller proportion of tumor cells showing nuclear staining (yellow arrows)

Fig 4: Time-dependent effect of vaspin on signaling pathways of MAP3/1, AKT, STAT3, PRKAA1, PKA, and expression of NFKB2 in coculture of Gc plus Tc and monoculture of Gc or Tc. Representative blots from five independent experiments are shown (A). The protein levels were shown as the ratio of phosphorylated protein to total protein for kinases, and the protein level in relation to actin for NFKB2 (B). Western blotting experiments were performed independently and repeated five times (n = 5). Data are plotted as the mean ± SEM. Significance between control and vaspin treatments is indicated by different letters (P < 0.05) from all five experiments.

Fig 5: (A) Schematic diagram of the alternative pathway of NF-kB (Reprinted from Lung Cancer, 89, 3, Dimitrakopoulos et al., 311–319, Variant of BCL3 gene is strongly associated with five-year survival of non-small-cell lung cancer patients, 2015, with permission from Elsevier). (B) Genomic positions of studied SNPs in NF-kB2 gene. (C) The study design outline. Abbreviations: BAFFR; tumor necrosis factor receptor superfamily member 13 C, CD40; CD40 molecule, TNF receptor superfamily member 5, LTßR; Lymphotoxin Beta Receptor (TNFR Superfamily, Member 3), RANK; Receptor Activator Of Nuclear Factor-Kappa B, NIK; NF-Kappa-Beta-Inducing Kinase, IKKa; I?B Kinase a, p100; nuclear factor NF-kappa-B p100 subunit, p52; nuclear factor NF-kappa-B p52 subunit, RELB; Transcription factor RelB, BCL3; B-Cell CLL/Lymphoma 3, FBXL15; F-Box and Leucine-Rich Repeat Protein 15, GBF1; Golgi Brefeldin A Resistant Guanine Nucleotide Exchange Factor 1, NF-kB2; nuclear factor-kappa B2, RPARP-AS1; RPARP antisense RNA 1.