Figure 6.

Myristoylation, subcellular distribution, and coimmunoprecipitation of Fyn with CD8-ζ after 2-hydroxymyristate treatment. Transfected cells expressing Fyn were preincubated overnight with 500 μM 2-hydroxymyristate, as described in Materials and Methods. (A) Cells were radiolabeled in the absence (−) or presence (+) of 500 μM 2-hydroxymyristate with Tran³⁵S-label, or ¹²⁵I-IC13, and analyzed directly (left) or subjected to homogenization and subcellular fractionation, followed immunoprecipitation (right). (B) COS-1 cells were cotransfected with cDNAs for Fyn and CD8-ζ, and incubated in the absence (−) or the presence (+) of 2-hydroxymyristate. Cells were lysed in digitonin buffer and coimmunoprecipitation of Fyn with CD8-ζ was analyzed using anti–CD8 antibody (+) or protein A/G⁺ agarose beads alone (−).