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. 2004 Mar;24(6):2455–2466. doi: 10.1128/MCB.24.6.2455-2466.2004

FIG. 7.

FIG. 7.

Lck is required for Tec tyrosine phosphorylation, and PLC-γ1 is tyrosine phosphorylated upon Tec overexpression. (A) Tec was transfected into Jurkat, JCaM1, Lck-reconstituted JCaM1, DT40, Lyn− Syk− DT40, and PLC-γ2− DT40 cells. Sixteen hours later, cells were lysed, and Tec was immunoprecipitated (i.p.) with Tec antiserum. Samples were Western blotted with antiphosphotyrosine MAb 4G10 or with Tec antiserum. (B) Jurkat cells were transfected with HA-PLC-γ1 in the presence or absence of Tec. Sixteen hours later, cells were harvested and stimulated with anti-TCR MAb C305 where indicated. PLC-γ1 was immunoprecipitated with anti-HA MAb and Western blotted with antiphosphotyrosine MAb 4G10 or with anti-PLC-γ1 MAb. Control Western blotting of whole-cell lysates (wcl) was done with 4G10 or Tec antiserum. (C) Jurkat cells were transfected with HA-PLC-γ1 in the presence or absence of HA-Tec. Sixteen hours later, cells were harvested and stimulated with anti-TCR MAb C305 where indicated. PLC-γ1 and Tec were immunoprecipitated with anti-HA MAb and Western blotted with rabbit anti-pY 783 PLC-γ1 or anti-HA MAb.