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Fig. 2 | BMC Veterinary Research

Fig. 2

From: Extracellular milieu grossly alters pathogen-specific immune response of mammary epithelial cells

Fig. 2

Influence of medium composition on pathogen or PAMP mediated NF-κB activation in pbMEC. a E. coli, but not S. aureus activates NF-κB in all media. pbMEC were transiently transfected with the NF-κB driven luciferase reporter gene and subsequently challenged for 24 h with 30 μg/ of heat killed pathogens in the media as indicated and the luciferase activity was measured from lysates (ordinate). Mean values (± S.E.M.) from five experiments (four biological replica) for SM0 and SM10, and two biological replica experiments for SM Milk. Each experiment was assayed in triplicate (*, p < 0.05; ***, p < 0.001 vs. control). b Only the E. coli challenge degrades IκBα. Degradation of IκBα was visualized in Western-blots after challenging the pbMEC in SM0 or SM10 with 30 μg/ml of heat killed E. coli or S. aureus 1027 particles. Thirty μg of cell lysates having been collected from unstimulated control cultures (c) or at the times as indicated after the challenge had been resolved in any slot of a 12 % SDS-gel and blotted onto nitrocellulose. The primary antibody was specific for IκBα. Data are representative for two independent experiments

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