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Fig. 8. CXCL12- and PMA-dependent regulation of ß2-chimaerin requires a functional C1 domain. (A) Jurkat T cells were transfected with a GFP-ß2-chimaerin C1 domain mutant (GFP-F215G). At 24 hours post-transfection, cells were sorted and GFP-positive cells tested for PMA-induced adhesion to sVCAM-1 as in Fig. 5. Data represent the mean ± s.d. of two independent experiments performed in triplicate. (B) Jurkat cells were transfected with GFP-ß2-chimaerin F215G and plated onto VCAM-1-coated slides, mounted on a 37°C plate and analyzed by confocal microscopy. CXCL12 (200 ng/ml) was added after the first frame and images are shown at the indicated times to illustrate cell spreading. Time-lapse confocal recording is shown in Movie 6 in supplementary material. (C) Jurkat T cells transfected with the indicated plasmids were sorted and GFP-positive cells were tested for CXCL12-induced adhesion to sVCAM-1 as in Fig. 5. Data represent the mean ± s.d. of two independent experiments performed in triplicate. Bar, 5 µm.
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