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Figure 4 | BMC Immunology

Figure 4

From: Interplay between the heterotrimeric G-protein subunits Gαq and Gαi2 sets the threshold for chemotaxis and TCR activation

Figure 4

Migration is inhibited by activation of SHP-1. (A) Jurkat TAg T cells were transfected with Gαq-specific siRNA (GNAQ1103), Gαi2-specific siRNA (GNAI-2-1050) or control siRNA (GNAQ1103 M3). 48 hours post-transfection cells were stimulated with CXCL12 for the indicated time periods and lysed in lysis buffer. SHP-1 immunoprecipitation was performed and immune complexes analysed by immunobloting with the indicated antibodies (n = 3). (B) Immunoblots from 3 independent experiments like in (A) were quantified by densiometric analysis. The graph show mean ± S.E.M. (C) Jurkat TAg T cells were tranfected with the phosphatase defective SHP-1 c/s (C453S) mutant or empty vector. 24 hours post-transfection cells were placed in Costar Transwell plates with or without CXCL12 (10 ng/ml) and migration analysed as above. Migration index represents fold increase in migration over basal migration in wells with vector tx cells (n = 3). (D) Activation of Gαi2 induces migration through a PI3K pathway whereas activation of Gαq inhibits migration through an Lck-SHP-1 pathway priming cell for activation through the TCR.

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