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. Author manuscript; available in PMC: 2010 Sep 17.
Published in final edited form as: Sci Signal. 2010 Aug 3;3(133):ra59. doi: 10.1126/scisignal.2000976

Fig. 3.

Fig. 3

Effects of the knockdown of Duox1 on TCR signaling in Jurkat cells. (A) Analysis of the abundances of Duox1, Duox2, and Nox2 proteins in Jurkat cell clones expressing NT or Duox1-specific (Duox1) shRNAs. The graph represents the average band density of Duox1 normalized to that in NT cells from three separate blots. *P < 0.05. WB, Western blotting. (B) Kinetics of TCR-induced oxidation of DCFDA in Jurkat cells stably transfected with NT (closed bars) or Duox1-specific (open bars) shRNA-expressing vectors. *P < 0.05. (C) TCR-induced activation of luciferase reporter plasmids driven by consensus sequences for AP-1 or NFAT in Jurkat cell clones expressing NT or Duox1-specific shRNAs. The data represent the mean fold increase in the amount of luciferase signal relative to that of unstimulated controls ± SEM from five experiments. *P < 0.05. (D and E) The kinetics of TCR-induced phosphorylation of ZAP-70 (Y319), ERK1 and ERK2, and Lck (Y394 and Y505) were determined by Western blotting analysis of whole-cell lysates from stable Jurkat clones expressing NT or Duox1-specific shRNAs with phosphospecific antibodies. Blots were then stripped and analyzed for total ZAP-70, ERK1 and ERK2, and Lck content to control for loading and to determine the effects of shRNAs.