Figure 6
Akt/mTOR/HIF1α pathway is indispensable for STING-mediated trained immunity. PM were pretreated with rapamycin (10 μM) for 1 h, and then the cells were trained with HKCA for 24 h and rested for 5 days. PM were restimulated with LPS or C. perfringens at an MOI of 5 for the indicated times. A, B TNF-α and IL-6 production were measured in the supernatants of PM response to LPS stimulation. C, D Phagocytosis and intracellular killing of C. perfringens was detected by CFU enumeration. E The cellular lysate was analyzed for p-mTOR, p-AKT, HIF1α, and β-actin by Western blotting. F–H The gray intensity value of proteins was calculated using ImageJ software. Data are shown as mean ± SEM. Data were pooled from 3 independent experiments. Statistical significance is indicated by *p < 0.05, **p < 0.01, and ***p < 0.001.