Modulating p56Lck in T-Cells by a Chimeric Peptide Comprising Two Functionally Different Motifs of Tip from Herpesvirus saimiri

Figure 6

Time-course assay for Lck in human PBMC after cell stimulation. (a) PBMC either untreated (lanes 1–4) or treated (lanes 5–8) for 1 h with hTip-CSKH were stimulated with PHA-P for 0, 15, 30, and 60 min and then were lysed. Extracts were subjected to SDS-PAGE and proteins transferred to PVDF membranes. WB was performed with Lck (upper panels) and P-Y (lower panels) monoclonal antibodies. Two bands were clearly defined which correspond to the low molecular weight (p56) and the high molecular weight band (p59), respectively. (b) Quantification of p56 and p59 bands showed an increase of the p59/p56 ratio. White bars represent nontreated cells and black bars hTip-CSKH-treated cells p59/p56 ratio at different time points. (c) Lck immunoprecipitation of PBMC extracts after stimulation with PHA-P or hTip-CSKH for 30 min. WB was performed with monoclonal antibodies against pTyr and Lck as indicated.

(a)

(b)

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