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. 2013 Jun 18;8(6):e66561. doi: 10.1371/journal.pone.0066561

Figure 1. Inhibitory effect of gliadin peptide 10-mer on IRAK1 phosphorylation and NF-kB activation.

Figure 1

CACO-2/TC7 cells, either unstimulated or stimulated with PT-Gly (1mg/ml), p10-mer (50 µg/ml), p10-mer (50 µg/ml) + PT-Gly (1mg/ml), were analyzed by Western blot for IRAK1 phosphorylation and NF-kB activation. (A) Phosphorylated levels of IRAK1 (p-IRAK1) were analyzed in whole cell extracts by Western blot with anti-phospho-IRAK1 antibodies; for control, the blotted membranes were stripped and reprobed with anti-IRAK-1 antibodies. Bound antibodies were visualized with HRP-conjugated IgG and immunoreactivity was assessed by ECL. (B) NF-kB activation was analyzed in whole cell extracts by Western blot with anti-phospho-NF-kB p65 Ser antibodies; for control, the blotted membranes were stripped and reprobed with anti-NF-kB p65 antibodies. Bound antibodies were visualized with HRP-conjugated IgG and immunoreactivity was assessed by ECL. Densitometric analysis was performed using ImageJ version 1.46 software and peaks were reproduced by reading the Western Blot bands. One example representative of 3 experiments.