MOESM4 of Streptococcus uberis strains isolated from the bovine mammary gland evade immune recognition by mammary epithelial cells, but not of macrophages

Additional file 4: S. uberis pretreatment of pbMEC did not change the immune response against a subsequent E. coli challenge. pbMEC were pretreated (primed) with 30 µg/mL heat-killed particles from S. uberis strain 0140J or 233 for one hour. Subsequently the cells were washed three times with PBS and cultivated in normal growth medium (0140J priming, 233 priming) or were challenged with 3 µg/mL heat-killed particles from E. coli strain 1303 (0140J priming + E. coli, 233 priming + E. coli) for another 1 h, 3 h or 24 h. To analyze the response against E. coli without priming pbMEC were cultivated one hour in normal growth medium, washed three times with PBS and were challenged with 3 µg/mL E. coli particles only for 1 h, 3 h or 24 h (E. coli). Cells were harvested at the end of the experiment and total RNA was prepared. TNF, CXCL8, NOS2A, and LAP mRNA concentrations were measured with RT-qPCR and expressed as multiples of the concentration from unstimulated controls. Data are from a single experiment, assayed in duplicate.