Fig. 8

Analysis of fresh and frozen explants to treatments with TLR-agonists and bacteria. A–B Evaluation of IL8 and IL6 release by ELISA. Teat explant were frozen at −80 ºC in 10% DMSO FBS. After thawing, explants were stimulated with LPS (1 µg/mL) or complete medium alone (control) for 24 h. Fresh counterparts from the same teat were used for comparison. C–D Relative IL-6 and IL-8 mRNA expression in explants as in A and B. E–G Analysis of cell populations by flow cytometry. Explants were treated with LPS (1 µg/mL), LTA (10 µg/mL), heat-killed E. coli or S. aureus. Complete medium alone was used as control. Percentages of leukocytes (E), granulocytes (F) and macrophages (G). Box-plots show data from 4 independent replicates (4 animals, 1 teat/animal). Data were analysed by Kruskal–Wallis test coupled to Dunn's multiple comparisons test. Asterisks indicate that values differ from control (^*P < 0.05)