FIG. 1.
Rat 2 cells expressing JCVE proteins and T125A and T125D mutant proteins. Whole-cell extracts were prepared from subconfluent cultures of R2CT, R2-JCVE, R2-T125A (four independent clones), and R2-T125D (two clones) cells, and proteins were immunoprecipitated with PAb 962 antibody. Coimmunoprecipitated proteins were separated in two 15% SDS-polyacrylamide gels (A and B), transferred to nitrocellulose membranes, and probed with a mixture of anti-T antibodies (PAb 962, PAb 2003, and PAb 2023) to enhance detection. Anti-rabbit IgG alkaline phosphatase-conjugated antibody was used as the secondary antibody, and protein bands were visualized using Nitro Blue Tetrazolium-5-bromo-4-chloro-3-indolylphosphate (NBT/BCIP) substrate buffer solution. The JCVE* and T125A* samples were treated with lambda protein phosphatase after the IP reaction. Protein molecular size standards (in kDa) are shown in lane M.