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. 2015 Feb 2;10(2):e0115116. doi: 10.1371/journal.pone.0115116

Figure 2. PMN cells from RA patients maintain constitutively activated ERK1/2.

Figure 2

(A) PMN cell lysates were prepared from RA patients and healthy donors and used to measure the activity of ERK by Western blot using antibodies specific for: phosphorylated or total ERK1/2. Concentrations of IL-8 (B), RANTES (C), IL-6 (D) and TNFα (E) were determined by ELISA from either the synovial fluid of RA patients or the serum of healthy donors. Error bars denote the SEM triplicate determinations of n = 19 RA samples and n = 6 controls, p values as shown. (F) Healthy PMN (n = 4) were isolated and cultured in the presence of either TNFα (1ng/ml), IL-6 (1ng/ml), or IL-17 (20ng/ml) for 24 and 48 hours before TREM1 analysis by flow cytometry (G) Healthy PMN (n = 4) were isolated and cultured in the presence of TNFα (0.01ug/ml to 0.1ug/ml) after the addition of TNFα blocking antibody for 24 and 48 hours before TREM1 analysis by flow cytometry. For both F and G error bars denote the SEM of duplicate readings of 4 treated primary healthy specimens. Asteriks denote p<0.05 against control untreated cells.