TNFα and IL-10 production by THP-1 macrophages stimulated with 2 mg/ml intact whey protein or whey hydrolysate in the presence or absence of celastrol.

<p>Celastrol significantly inhibited both TNFα and IL-10 induced by intact whey protein and whey hydrolysates, demonstrating NF-κB dependency of the cytokine production. Significant differences were determined by using paired T tests. Significant differences were indicated by *.</p

NF-kB/AP-1 expression in HEK reporter cells carrying individual TLRs after stimulation with intact whey protein or hydrolysate.

<p>Intact whey protein induced signaling of TLR 2, 4, and 7, but this pattern of TLR activation by whey changed after hydrolysis. The TLR activating capacity of whey hydrolysates decreased with increasing level of hydrolysis. Significant differences compared to the negative control were determined by using the Kruskal-Wallis test followed by the Dunn’s test and indicated by *.</p

NF-kB/AP-1 expression in THP-1-MD2-CD14 and THP-1-MyD88 deficient reporter cells after stimulation with intact whey and casein proteins or hydrolysates.

<p>Intact whey and the two most mildly hydrolyzed whey hydrolysates induced TLR signaling, while casein samples had no effect (left panel). These effects were TLR dependent, since no activation was observed in the MyD88 deficient cell line after stimulation (right panel). Significant differences compared to the negative control were determined by using the Kruskal-Wallis test followed by the Dunn’s test and indicated by *.</p

Human primary PBMCs produced cytokines after stimulation with intact whey and casein proteins or hydrolysates.

<p>Whey samples induced TNFα and IL-10 in a degree of hydrolysis dependent way. Not many effects were observed for IL-8 (upper panel). Effects induced by casein samples was less compared to the whey samples. No TNFα and IL-10 were produced after stimulation with intact casein or its hydrolysates. IL-8 production was increased by some casein samples (lower panel). Significant differences compared to the negative control were determined by using the Kruskal-Wallis test followed by the Dunn’s test and indicated by *.</p

Cell densities and ligands used in the different reporter cell line assays.

<p>Cell densities and ligands used in the different reporter cell line assays.</p

Overview of the characteristics of the studied hydrolysates.

<p>Overview of the characteristics of the studied hydrolysates.</p

NF-kB/AP-1 expression in HEK reporter cells carrying individual TLRs after simultaneous stimulation with its relevant ligand and intact casein protein or hydrolysate.

<p>Casein samples were very efficacious in inhibiting TLR signaling. Intact casein strongly inhibited TLR 9. Hydrolysis of casein did change the magnitude of inhibition and the type of TLRs that were inhibited. Significant differences were determined by using the Kruskal-Wallis test followed by the Dunn’s test. Significant differences compared to the negative control were indicated by *, significant differences compared to the positive control were indicated by #.</p

NF-kB/AP-1 expression in HEK reporter cells carrying individual TLRs after stimulation with intact casein protein or hydrolysate.

<p>Intact casein stimulated TLR2, 3, 4, 5 and 9. The TLR activating capacity was completely lost after more extensive hydrolysis. Significant differences compared to the negative control were determined by using the Kruskal-Wallis test followed by the Dunn’s test and indicated by *.</p