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The EMBO Journal (2004) 23, 2577–2585, doi:10.1038/sj.emboj.7600268 Published online 10 June 2004 |
| Figure 3 |
| Negative feedback loop in T-cell activation through MAPK-catalyzed threonine phosphorylation of LAT |
| Satoshi Matsuda, Yoshihiro Miwa, Yasuko Hirata, Akiko Minowa, Junko Tanaka, Eisuke Nishida and Shigeo Koyasu |
mAb 2Ad2A2 for the indicated times. Immunoprecipitated LAT was subjected to immunoblot with 4G10 or the mAb against phospho-Thr-Pro. (B) Jurkat cells were stimulated with 2Ad2A2 for the indicated times (
CD3) or with pervanadate for 10 min (PV). The lysates were subjected to immunoblot analysis with an mAb against phospho-ERK (Cell Signaling) (p-ERK2) and the anti-ERK2 antibody (ERK2). The lysates were also subjected to immunoprecipitation with anti-JNK1 antibody followed by the kinase assay using ATF2 as a substrate (p-ATF2). Immunoprecipitation of JNK1 was confirmed with immunoblot (JNK). (C) Jurkat cells were stimulated with (+) or without (-) 2Ad2A2 for 3 min in the presence or absence of 50 
M PD98059. Immunoprecipitated LAT was subjected to immunoblot with the mAb against phospho-Thr-Pro (upper panel) or 4G10 (an mAb against phospho-Tyr) (lower panel).
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