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. 2011 Nov 28;11(1):104–110. doi: 10.1111/j.1474-9726.2011.00759.x

Figure 3.

Figure 3

Flagellin-stimulated monocytes from the elderly show increased phosphorylation of p38 and ERK MAPK. (A) immunoblot of p38 phosphorylation in a representative result of young and old individuals after stimulation with the toll-like receptor (TLR)5 ligand flagellin in monocytes. Phosphorylation levels at 5, 10, and 30 min are compared with unstimulated control samples. (B) Densitometry of immunoblot of p38 and phospho-p38 in monocytes from 21 pairs of younger and older subjects after stimulation with flagellin for 10 min. Densitometry shows the means ± SEM of the ratio of total p38 and phospho-p38 to β-actin, and of phospho-p38 to total p38. Asterisks indicate statistical significance between younger and older cohort (***P < 0.001). (C) Correlation of TLR5 expression with p38 phosphorylation after stimulation with flagellin (n = 42, Pearson r = 0.3165, P = 0.0438). (D) Densitometry of immunoblot of ERK in monocytes from 21 pairs of younger and older subjects after stimulation with flagellin for 10 min. Densitometry shows the means ± SEM of the ratio of total ERK and phospho-ERK to β-actin, and of phospho-ERK to total ERK. Asterisks indicate statistical significance between younger and older cohort (**P < 0.01). (E) Correlation of ERK phosphorylation with p38 phosphorylation after stimulation with flagellin (n = 42, Pearson r = 0.5336, P = 0.0004). (F) Monocytes from older (n = 20) and young (n = 20) adults were stimulated with TLR5 ligand flagellin for 20 h. IL-1β, IL-6, and MIP-3α were measured from culture supernatants by ELISA. Data shown are the means ± SEM; asterisks indicate statistical significance between young and older cohort (**P < 0.01).