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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. 4.

Fig. 4

Effects of the increased abundance of Duox1 on TCR signaling in Jurkat cells. (A) The Duox1-knockdown Jurkat cell clone was transfected with an empty vector or with a plasmid encoding HA-tagged Duox1. TCR-induced phosphorylation of ZAP-70 (Y319), ERK1 and ERK2, and Lck (Y394 and Y505) was detected after 5 min of stimulation by Western blotting analysis of whole-cell lysates with phosphospecific antibodies. Blots were then stripped and analyzed for total PLC-γ1, ERK1 and ERK2, and Lck content to control for loading and to determine the effects of the increased abundance of Duox1. (B) Jurkat cells were transiently transfected with an empty vector or with a plasmid encoding HA-tagged Duox1. TCR-induced phosphorylation of PLC-γ1, ZAP-70 (Y319), and ERK1 and ERK2 was analyzed as described for (A). Blots were then stripped and analyzed for total PLC-γ1, ZAP-70, and γ-tubulin content to control for loading. (C) Activation of ERK1 and ERK2 was measured in Jurkat cells transiently transfected with empty vector, plasmid encoding HA-tagged Duox1, or plasmid encoding HA-tagged Duox2. HA-tagged proteins were immunoprecipitated from their respective lysates and analyzed by Western blotting with antibodies against Duox1 or Duox2.