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Fig. 6 | Arthritis Research & Therapy

Fig. 6

From: Dysregulation of RasGRP1 in rheumatoid arthritis and modulation of RasGRP3 as a biomarker of TNFα inhibitors

Fig. 6

Functional effects of TNFα in B and T cells. a Cell proliferation was evaluated with (3H) thymidine incorporation by B and T cells with or without TNFα (n = 3). b Analysis of mitogen-activated protein kinase (MAPK) phosphorylation in B and T cells with or without TNFα for 48 hours was performed with a human phospho-kinase array kit to measure Lyn, Lck, PLCγ1, MEK, ERK and RSK phosphorylation (n = 2). The dotted line represents the control condition without TNFα. c Western blot analysis was performed to measure ERK1/2 and phospho-ERK1/2 protein expressions in T cells from five healthy controls (HC). Cells were cultured with or without TNFα for 48 hours and with or without antibodies targeting TNFR1 or TNFR2. Histogram represents ratio of phosphorylated ERK1/2 to total ERK. A value of 1 was arbitrarily assigned to control conditions (without TNFα treatment) to which glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was reported and expressed as fold change. d After incubation with 1 ng/ml of TNFα for 48 hours, cells were stimulated with 50nM Bryostatin-1 for 5 hours. CD69 expression in T cells (CD3) was assessed using flow cytometry (one representative result of three independent experiments is presented). Mean ± standard error of the mean were compared using Student’s t test or one-way analysis of variance followed by Dunnett post-hoc test. *p <0.05; **p <0.01

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