TOM1L Is Involved in a Novel Signaling Pathway Important for the IL-2 Production in Jurkat T Cells Stimulated by CD3/CD28 CoLigation
Figure 2
(a) 293T cells transfected with plasmids encoding Fyn alone or together with Myc-tagged full length TOM1L. Fyn and TOM1L proteins were isolated by immunoprecipitation using anti-Fyn, or anti-Myc antibodies and assayed by SDS−PAGE and immunoblotting as described in material and methods. Immunoblots were analyzed with primary anti-phosphotyrosine (anti-PTyr), anti-Fyn or anti-Myc –antibodies, respectively. (b) 293T cells transfected with plasmids encoding Lck alone or together with Myc-tagged full length TOM1L. Lck and TOM1L proteins were isolated by immunoprecipitation using anti-Lck or anti-Myc antibodies and assayed by SDS−PAGE and immunoblotting as described in material and methods. (c) Kinase assay: Anti-Fyn immunoprecipitates were incubated with 5 Ci of (-32P) ATP and 2 mM (Val5)-angiotensin II as an exogenous substrate in PIPES kinase buffer for 1, 3, and 5 minutes at room temperature.
