Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Intensity and duration of TCR signaling is limited by p38 phosphorylation of ZAP-70 T293 and destabilization of the signalosome

doi: 10.1073/pnas.1713301115

Figure Lengend Snippet: Phosphorylation of ZAP-70T293 diminishes recruitment of ZAP-70 to the TCR and destabilizes the signaling complex. (A) ZAP-70–deficient P116 cells expressing WT ZAP-70 or ZAP-70T293A were activated with cross-linked anti-CD3 and cell lysates prepared at the indicated times were immunoprecipitated with anti–TCR-ζ. The immunoprecipitates were resolved by SDS/PAGE and immunoblotted with antibodies against the indicated proteins. The data are representative of three independent experiments. (B, Upper) Representative images illustrating the time course of phosphorylated ZAP-70 clustering in activated Jurkat cells. The images shown here were taken from different staining experiments. (B, Lower) Representative images illustrating the time course of phosphorylated LAT clustering in activated Jurkat cells. The images shown in the Upper and Lower panels were taken from different staining experiments. (C) Quantification of the area of pZAP-70 and pLAT clusters. Error bars show SEM. For WT ZAP-70 cells, 3 min n = 51, 6 min n = 55, 9 min n = 52, 12 min n = 55, and 15 min n = 60. For ZAP-70T293A cells, 3 min n = 55, 6 min n = 50, 9 min n = 52, 12 min n = 53, and 15 min n = 52, taken from three experiments. AU, arbitrary units.

Article Snippet: Agarose-conjugated anti–TCR-ζ was from Santa Cruz.

Techniques: Expressing, Immunoprecipitation, SDS Page, Staining