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. Author manuscript; available in PMC: 2007 Mar 22.
Published in final edited form as: Cell Immunol. 2006 Nov 2;242(2):80–90. doi: 10.1016/j.cellimm.2006.09.008

Figure 6. Activation of Lck is compromised in anti-CD3 treated memory CD4 T cells.

Figure 6

A–B, Naive (N) and memory (M) DO11.10 T cells were stimulated 2 min with OVA, SEB, anti-CD3 or left unstimulated (Unstim.). Lck protein was immunoprecipitated (ip) from the T cell lysates and (A) resolved on 8% SDS-PAGE gels and immunoblotted (blot) with anti-phosphotyrosine and anti-Lck Abs, or (B) used in tyrosine kinase assays. Phosphorylation values were determined by converting Absorbance measurements using a standard curve derived from supplied soluble phosphopeptide (ng/mL). C–D, Naive (○, ●) and memory (▽, ▼) DO11.10 T cells were stimulated with anti-CD3 (open) or SEB (filled) for the indicated lengths of time (min). Lck protein was immunoprecipitated from the T cell lysates and (C) resolved on 8% SDS-PAGE gels and immunoblotted as above, or (D) used in tyrosine kinase assays as above. Densitometry was performed on the immunoblots and the relative levels of phosphoprotein are expressed as a ratio relative to the total amount of precipitated protein. Data are representative of at least two independent experiments.