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. Author manuscript; available in PMC: 2015 Feb 1.
Published in final edited form as: Mol Immunol. 2013 Oct 12;57(2):10.1016/j.molimm.2013.09.002. doi: 10.1016/j.molimm.2013.09.002

Figure 5.

Figure 5

Flagellin pretreatment differentially affects early TCR-induced signaling in HuT78 cells. (A) HuT78 T cells were pretreated for 24 hours with or without 250 ng/ml flagellin. The cells were then stimulated with 10 μg/ml anti-CD3 (OKT3) for various times and lysed. The proteins were separated by PAGE. The phosphorylation of Lck, (top panel) LAT tyrosine 191 (second panel), ERK1/2 (third panel), Akt (fourth panel), and the expression of actin (bottom panel) were examined by immunoblotting. (B) The results were normalized to actin and the maximal level of activation in the individual experiment. The results are an average of three to five independent experiments ± SEM. The data was statistically compared (* = p<0.05 and ** = p<0.01) using Student's t test. (C) HuT78 T cells were pretreated for 24 hours with or without 250 ng/mL flagellin. The cells were then incubated with the calcium sensitive dye Fluor-4. The TCR-induced influx of calcium into the T cells was assessed by flow cytometry with Ionomycin serving as a positive control. The results are representative of three separate experiments.