2.5. Bone Marrow Precursor Cell Treatments

Endotoxin-free ovalbumin (EndoGrade-ovalbumin) supplemented with 1 ng/mL lipopolysaccharide (LPS) at the final concentration was used as endotoxin-containing model pathogen. To investigate the effects of endotoxin-containing pathogen on Akt/IKKα/β activation and the endosomal translocation of Sec61, BMPC was incubated with endotoxin-containing model pathogen or endotoxin-free ovalbumin at the final concentration of 50 μg/mL. To inhibit related kinase activities, BMPC was pretreated with wortmannin (5 μM), Bay11-7082 (5 μM) or siRNA transfection prior to the incubation with endotoxin-containing model pathogen (50 μg/mL). For cytometric analyses of cross-presented OVA, the BMPC was conferred with endotoxin-containing model pathogen for 7 h.