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. 2016 Jun 9;11(6):e0157175. doi: 10.1371/journal.pone.0157175

Fig 4. Prior exposure to IL-12 alters select TCR-induced signaling pathways in human activated CD4 T cells.

Fig 4

Human activated CD4 T cells were (A and B) treated with or without IL-12 (50 ng/mL) for different times or (C and D) incubated with or without 50 ng/mL of IL-12 for 6 h and then stimulated with anti-TCR (anti-CD3 clone OKT3) and anti-CD4 (clone RPA-T4) antibodies for various times. The phosphorylation of signaling molecules was determined in whole cell lysates by immunoblotting. Results were normalized to GAPDH and the maximal level of activation of no cytokine cells. Data are shown as (A and C) representative blots from different donors and (B and D) mean ± SEM of normalized results of three to six separate donors. The loading controls shown for each figure correspond to at least one of the blots shown but for the quantification each blot was quantified with its respective control. Data were analyzed with two-tail, unpaired Student’s t test. *p<0.05; **p<0.01; ***p<0.001; n.s. = not significant.