EGCG prevents PCB-126-induced endothelial cell inflammation via epigenetic modifications of NF-κB target genes in human endothelial cells Dandan Liu,

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EGCG prevents PCB-126-induced endothelial cell inflammation via epigenetic modifications of NF-κB target genes in human endothelial cells Dandan Liu, Jordan T. Perkins, Bernhard Hennig Journal of Nutritional Biochemistry Volume 28, Pages 164-170 (February 2016) DOI: 10.1016/j.jnutbio.2015.10.003 Copyright © 2015 Elsevier Inc. Terms and Conditions

Fig. 1 EGCG decreases PCB-induced NF-κB activation. (A) EGCG decreased PCB-126-induced p65 expression. Cellular p65 mRNA level was measured in endothelial cells which were pretreated with 15 or 30 μM of EGCG (E15, E30) for 8 h and then treated in fresh medium with 0.03 nM of PCB 126 for 16 h. Values are means±SD with n=3. ⁎Significantly increased compared to the DMSO control. #Significantly decreased compared to the PCB treatment group. (B) Protein levels of p65, IKK-α/β, IκB-α and IκB-β were measured by immunoblotting. Actin was used as the loading control. (C) EGCG decreased PCB-126-mediated p65 nuclear translocation. The intracellular localization of endogenous p65 was analyzed by immunocytochemical staining using anti-p65 (green), and the nucleus was visualized by DAPI staining (blue). Journal of Nutritional Biochemistry 2016 28, 164-170DOI: (10.1016/j.jnutbio.2015.10.003) Copyright © 2015 Elsevier Inc. Terms and Conditions

Fig. 2 EGCG attenuates PCB-126-induced inflammatory gene expression. The mRNA level of inflammatory genes was analyzed using qRT-PCR. Endothelial cells were pretreated with 15 or 30 μM of EGCG for 8 h and followed by treatment in fresh medium with 0.03 nM of PCB 126 for 16 h. Values are the means±SD with n=3. ⁎Significantly increased compared to the DMSO control. ^Significantly decreased compared to the DMSO control. #Significantly decreased compared to the PCB treatment group. Journal of Nutritional Biochemistry 2016 28, 164-170DOI: (10.1016/j.jnutbio.2015.10.003) Copyright © 2015 Elsevier Inc. Terms and Conditions

Fig. 3 EGCG reverses PCB-126-induced chromatin binding of acetylation enzymes. The recruitment of p65, p300 and HDACs 1–3 was modulated by EGCG in the promoters of IL-6 (A), CRP (B), ICAM-1 (C), VCAM-1 (D), IL-1α (E), IL-1β (F) and GAPDH (G). Sheared chromatin from whole-cell lysates was immunoprecipitated with antibodies against the indicated proteins. IgG was used as a negative control. The GAPDH promoter was used as control locus. Values are means±SD with n=3. ⁎Significantly increased compared to the DMSO control. ^Significantly decreased compared to the DMSO control. #Significantly decreased compared to the PCB treatment group. Journal of Nutritional Biochemistry 2016 28, 164-170DOI: (10.1016/j.jnutbio.2015.10.003) Copyright © 2015 Elsevier Inc. Terms and Conditions

Fig. 4 EGCG induces H3 hypoacetylation as a result of HAT/HDAC binding exchange. The H3 acetylation was enhanced by PCB 126 and decreased by the pretreatment with EGCG in the NF-κB-regulated genes. H3 is the control for nucleosomal occupancy. Values are means±SD with n=3. ⁎Significantly increased compared to the DMSO control. ^Significantly decreased compared to the DMSO control. #Significantly decreased compared to the PCB treatment group. Journal of Nutritional Biochemistry 2016 28, 164-170DOI: (10.1016/j.jnutbio.2015.10.003) Copyright © 2015 Elsevier Inc. Terms and Conditions