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The abundance of phosphorylated form was always related to the amount of non-phosphorylated protein assessed on the same membrane after incubation with stripping buffer (Pierce). The activation of the particular transduction pathway was presented as ratio between phosphorylated form and total abundance of the protein, and this ratio in average WT placebo treated animal was arbitrarily set as 1. The experimental procedures were performed on the extensively studied and well characterized C57BL/6JIL-6-/-TMKopf learn more strain [20, 21, 22?and?23], and results were compared to the respective wild type C57BL/6J animals. The genotypes were assessed as previously described using standard PCR of DNA isolated from mouse tails [22]. Results were presented as the mean �� standard deviation (SD). The student t-test was used for statistical analysis. A p value less than 0.05 was considered statistically Fleroxacin significant. After placebo injection STAT3 phosphorylation is significantly more pronounced in WT animals as compared to IL-6 KO mice (p buy CHIR-99021 1B). After 24 hours the extent of STAT 3 phosphorylation returns to the levels close to the initial state both in WT and IL-6 KO. The assessed knockout did not influence the activation of MEK/ERK pathway (the extent of ERK 1 and ERK2 phosphorylation in relation to the total ERK 1 and ERK2 protein) after placebo injection. Isoproterenol delivery caused rapid and potent (approximately 10 fold) (p