Table 1.

Diffusion and Single-Molecule Centroid Positions Relative to CD2-mRFP Clusters

Construct	Din (µm2/s)	Dout (µm2/s)	Din/Dout	p value (Din/Dout)	Nin/Nout	p value (Nin/Nout)
LAT	0.18	0.30	0.59 ± 0.06	< 0.005	1.80 ± 0.31	< 0.05
LAT(Y-F)	0.66	0.55	1.23 ± 0.26	–	0.89 ± 0.12	–
LAT(C-S)	0.14	0.43	0.32 ± 0.11	<0.005	2.1 ± 0.21	<0.01
Lck	0.26	0.46	0.55 ± 0.11	<0.10	1.45 ± 0.28	–
Lck10	1.08	1.14	0.95 ± 0.04	–	0.89 ± 0.11	–
CD45	N.A.	N.A.	N.A.	N.A.	0.64 ± 0.07	<0.01

The centroids of images of single GFP-tagged proteins were overlaid onto the CD2-mRFP image, as described in the Experimental Procedures. D_in refers to the mean diffusion coefficients for trajectory segments overlapping a CD2 zone for five or more consecutive frames, and D_out indicates the mean diffusion coefficient for trajectory segments lying outside of CD2 zones for five or more consecutive frames. CD45 is listed as N.A. (not analyzable), since the number of trajectories that had overlapping centroids with CD2 zones for five consecutive frames was >10-fold lower than LAT-GFP, and some cells did not have any CD45 trajectories that fit this criteria. N_in/N_out refer to the ratio of centroid coordinates that are found inside or outside of CD2 clusters, normalized to the fraction of the area occupied by CD2 clusters. A value of one would be expected for a random distribution. Four to nine cells were analyzed for each construct. Error estimates represent the SEM for the different cells analyzed. p values indicate the significance of the deviation from a value of 1.