Journal: Veterinary Research
Article Title: Regulation of tracheal antimicrobial peptide gene expression in airway epithelial cells of cattle
Figure Lengend Snippet: Western analysis of NF-kB p65 in nuclear and cytoplasmic extracts of bTEC stimulated with LPS, Pam3CSK4 or IL-17A. NF-kB p65 was localized to the cytoplasm of non-stimulated cells (NS), but showed translocation to the nucleus following treatment of the cells with either LPS (panel A ), Pam3CSK4 (Pam, panel B ) or IL-17A (panel C ). However, pre-treating the cells with the NF-κB inhibitor CAPE (10 μM) prior to exposure to agonists fully abrogated nuclear translocation of NF-kB p65 in the studies involving LPS and Pam3CSK4 (panels A and B ), and partially in the studies involving IL-17A (panel C ). Pos control: positive control, NF-kB p65. The findings were similar when the experiments were repeated using cells from different animals.
Article Snippet: Triplicate cell cultures supplemented with DMEM/F12 without serum were unstimulated, or stimulated with 0.1 μg/mL LPS (Sigma Aldrich, MO, USA, L9143), 1 μg/mL Pam3CSK4 (Invivogen, San Diego, CA, USA, t1rl-pms), or 316 ng/mL IL-17A (Kingfisher Biotech, St. Paul, MN, USA, RP0056B-005) for 16 h. The doses and time were optimized in a previous study [ ].
Techniques: Western Blot, Translocation Assay, Positive Control