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. 2020 Jun 13;21(12):4225. doi: 10.3390/ijms21124225

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© 2020 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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Cell-permeable NBD peptide inhibits NF-κB activation, inflammation and oxidative stress in vitro. (A) Immunoprecipitation of NEMO with IKKα/β subunits in mesangial cells treated with NBD peptide (50 µM, 90 min) before stimulation with high-glucose and TNFα (HGG+ TNFα, 2 h); (B) western blot analysis of nuclear p65 protein (loading control, histone H3) in mesangial cells; (C) real-time PCR analysis of proinflammatory genes (normalized to 18S) in mesangial cells pretreated with NBD and Mut peptides prior to stimulation (HG + TNFα, 24 h); (D) measurement of NADPH oxidase activity (expressed as relative light units (RLU) per mg) in macrophage cell lysates after 3 h of stimulation with lipopolysaccharide (LPS) in the presence or absence of NBD and Mut peptides. Results presented as individual data points and mean ± SD of 4 independent experiments. *—p < 0.05 vs. basal; #—p < 0.05 vs. stimulus; $—p < 0.05 vs. Mut.

Cell-permeable NBD peptide inhibits NF-κB activation, inflammation and oxidative stress in vitro. (A) Immunoprecipitation of NEMO with IKKα/β subunits in mesangial cells treated with NBD peptide (50 µM, 90 min) before stimulation with high-glucose and TNFα (HGG+ TNFα, 2 h); (B) western blot analysis of nuclear p65 protein (loading control, histone H3) in mesangial cells; (C) real-time PCR analysis of proinflammatory genes (normalized to 18S) in mesangial cells pretreated with NBD and Mut peptides prior to stimulation (HG + TNFα, 24 h); (D) measurement of NADPH oxidase activity (expressed as relative light units (RLU) per mg) in macrophage cell lysates after 3 h of stimulation with lipopolysaccharide (LPS) in the presence or absence of NBD and Mut peptides. Results presented as individual data points and mean ± SD of 4 independent experiments. *—p < 0.05 vs. basal; #—p < 0.05 vs. stimulus; $—p < 0.05 vs. Mut.