Fig. 3
BMMs from IRF5−/− mice demonstrate reduced PRR-induced PDK1 activation and PI3K/PDK1-dependent phagocytosis. a, b BMMs from IRF5+/+, IRF5+/− or IRF5−/− mice were left untreated or treated for 48 h with 0.1 μg/ml lipid A (mature). a Cells were then co-cultured with S. Typhimurium-GFP, or FITC-labeled E. coli or S. aureus bioparticles for 20 min, and intracellular uptake was examined by flow cytometry (mean fluorescence intensity (MFI) + SEM) (3 replicates, representative of 2 independent experiments). b Cells were treated with lipid A for 20 min and examined for phosphorylated-PDK1 by Western blot. PDK1 and GAPDH served as loading controls. c BMMs were treated for 48 h with 0.1 μg/ml lipid A, then pre-treated with a PDK1 inhibitor (GSK) or wortmannin for 1 h and then co-cultured with bacteria or bacterial particles as per (a). MFI + SEM (3 replicates). Significance is compared to IRF5+/+ macrophages for each respective treatment condition or as indicated. Inh, inhibitors. *p < 0.05; **p < 0.01; ***p < 0.001; †p < 1 × 10−4; ††p < 1 × 10−5
