Figure 4. Tyrosine Phosphorylation of p58 and Phosphorylation-Dependent Association with HCP Reconstituted in Nonhematopoietic.

Cells 3T3E cells (5 × 10⁶) were infected for 6 hr with 5 pfu/cell of recombinant vaccinia viruses in combination with the appropriate dose of the control virus (Vac–c) to normalize the infections to 15 pfu/cell. The recombinant viruses express lyn tyrosine kinase (Vac–lyn), HCP (Vac–HCP), p58-cl6 (Vac–6). Following lysis with Lysis-L buffer, all samples were subjected to immunoprecipitation with GL183. The samples were divided into two lanes of the same gel such that, following transfer to a membrane, they were probed in parallel with anti-phosphotyrosine (αptyr) or anti-HCP (αHCP). The amount of p58 in each sample was determined by reprobing with αcyt6 the membrane that had been first probed with αHCP. The position corresponding to the migration of p58 is indicated on the right, along with those of molecular mass markers.