Integrin α₃β₁-binding peptide LXY2 differentially regulates cytokine secretion due to TLR2 and TLR4 stimulation. (A) ELISA of IL-6, TNF-α, and IL-10 in supernatants of neutrophils, isolated by negative selection, from BM of wild-type C57Bl/6 mice. Cells (10⁶ cells/300 μL) were incubated for 24 hours with Pam₃CSK₄ (100 ng/mL) with and without α₃β₁-binding peptide LXY2 (1 μM). Supernatants were tested in duplicate or triplicate. The results from 3 independent experiments are expressed as mean ± SD. *P < .05 (Mann-Whitney test). (B) Left: Representative western blot images of FAK phosphorylation kinetics in neutrophils (600 000 cells/100 μL) from C57BL/6 mice after Pam₃CSK₄ (10 μg/mL) stimulation ± LXY2 (100 μM) over indicated periods. Total cell lysates were analyzed by western blotting to determine the extent of FAK phosphorylation at Tyr³⁹⁷. Right: Densitometric analysis of FAK phosphorylation in WT neutrophils from C57BL/6 mice. Levels of phospho-FAK were normalized to total FAK at each time point and expressed as mean fold increase compared with control (time = 0 minutes). Data are expressed as mean ± SEM from 3 independent experiments. *P < .05 (2-way repeated measures ANOVA with Bonferroni posttest).