Figure 1.
Figure 1. SKAP55 is recruited into microclusters by SLP-76 and ADAP. (A) Immunofluorescent staining for endogenous SKAP55 in J14.SY cells fixed 5 min after plating on anti-CD3 coated, BSA-blocked stimulatory coverslips. (B) J14.SY cells expressing either SKAP55.TRT (top) or TRT (bottom) were stimulated on coverslips and imaged for 5 min. MOT projections depict the trajectories of microclusters over time (n = 20). (C) MOT images of J14 cells coexpressing SKAP55.mRFP1 and either mYFP (top) or SLP-76.mYFP (bottom) (n = 3). (D) MOT images of ADAP-deficient JDAP cells coexpressing SKAP55.3×Flag.mCFP, SLP-76.YFP, and either 3×Flag.TRT (top) or TRT.3×Flag.TRT.ADAP-120 (bottom) (n = 4). (E) Kymographs depicting the lateral movements of SLP-76, SKAP55, and ADAP microclusters along a narrow region of interest spanning the center of the cell. Bars: (A–D) 10 µm; (E) 5 µm × 60 s.

SKAP55 is recruited into microclusters by SLP-76 and ADAP. (A) Immunofluorescent staining for endogenous SKAP55 in J14.SY cells fixed 5 min after plating on anti-CD3 coated, BSA-blocked stimulatory coverslips. (B) J14.SY cells expressing either SKAP55.TRT (top) or TRT (bottom) were stimulated on coverslips and imaged for 5 min. MOT projections depict the trajectories of microclusters over time (n = 20). (C) MOT images of J14 cells coexpressing SKAP55.mRFP1 and either mYFP (top) or SLP-76.mYFP (bottom) (n = 3). (D) MOT images of ADAP-deficient JDAP cells coexpressing SKAP55.3×Flag.mCFP, SLP-76.YFP, and either 3×Flag.TRT (top) or TRT.3×Flag.TRT.ADAP-120 (bottom) (n = 4). (E) Kymographs depicting the lateral movements of SLP-76, SKAP55, and ADAP microclusters along a narrow region of interest spanning the center of the cell. Bars: (A–D) 10 µm; (E) 5 µm × 60 s.

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