The p110𝛼 and p110𝛽 Isoforms of Class I Phosphatidylinositol 3-Kinase Are Involved in Toll-Like Receptor 5 Signaling in Epithelial Cells

Figure 5

Effect of p110$\alpha$- and p110$\beta$-targeted shRNA on IL-8 production in TLR5-transfected, flagellin-stimulated HEK293T cells. Cells were transfected with TLR5 and a pSuper derivative or with pSuper alone and then stimulated with 500 ng/ml flagellin for 6 hours. IL-8 concentrations were determined by ELISA. pSp110-$\alpha$1 and -$\alpha$2 target 2 separate sites of p110$\alpha$, while pSp110-$\beta$1 and -$\beta$2 target two separate sites of p110$\beta$. Results show fold increase versus baseline (IL-8 amounts measured in flagellin stimulated cells which were not transfected with TLR5). The percentages above the columns denote the amount of knockdown of either p110$\alpha$ or p110$\beta$ mRNA as determined by quantitative RT-PCR; results were compiled from at least 7 samples from a minimum of 3 separate experiments. All four knockdowns significantly reduced IL-8 production ($P<.001$, ANOVA).