Figure 4

Western blot analysis of IKK-α, IkBα, and NFkB in serous papillary ovarian carcinoma. Representative protein profiles (A) of cytosolic and nuclear extracts (70 μg protein) pooled from 10 samples/group. (B–G). Extracts obtained from individual rats were used for densitometric analysis of the protein levels following normalization to β-actin or LaminB1. The results are expressed as the means ± SD (n = 10 animals/group). ^aP < 0.05 vs. OC; ^bP < 0.05 vs. OC + Mel; and ^cP < 0.05 vs. OC + EtOH. (H) The canonical pathway that leads to the activation of NFkB after a ligand binds to its specific receptor. Internal signaling results in the phosphorylation of IkBα by the IKK complex, leading to the ubiquitylation and degradation of NFkB via proteasome. The NFkB p65/p50 complex translocates to the nucleus to initiate the transactivation of target genes related to the inflammatory process. Mel induces the downregulation of IKK-α, IkBα, and NFkB p65/p50, whereas EtOH consumption upregulates IKK-α and NFkB p65. (I-L) Merged images of NFkB p65 immunofluorescence and DAPI nuclear staining in the OC (I), OC + Mel (J), OC + EtOH (K), and OC + EtOH + Mel groups (L); details of cytosolic/nuclear staining (Alexafluor®488, Bar = 10 μm). EtOH: ethanol; mel: melatonin; IKK-α: inhibitor of NFkB kinase subunit alpha; IkBα: inhibitor of NFkB subunit alpha; NFkB p65: NFkB subunit p65 (RelA); NFkB p50: NFkB subunit p50.