To investigate the roles of various hematopoietic cell-specific adapter proteins in T cell receptor (TCR)-signaling leading to nuclear factor of activated T cell (NF-AT) and nuclear factor of κB (NF-κB) activation, we reconstituted TCR-signaling with CD8/ζ, various protein tyrosine kinases (PTKs), and adapter proteins in a non-lymphoid cell line, 293T. We show that SLP-76 and BLNK, but not LAT, effectively co-operated with Syk and Tec family PTKs to activate NF-AT and NF-κB. We also show that Tec family PTKs enhanced endogenous phospholipase C (PLC)-γ1 phosphorylation induced by CD8/ζ and Syk in 293T cells. These results imply that PLC-γ1 may play a critical role in a hematopoietic cell-specific adapter protein-mediated NF-AT and NF-κB activation in a non-lymphoid cell.
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