FIGURE 1:

The C-terminal fragment containing the KD is required for PKCθ IS retention. (A) The indicated D10 T cell lines were stimulated with CH12 cells loaded with no antigen (No Ag) or 250 μg/ml conalbumin (+Antigen). PKCθ-GFP constructs were visualized by digital deconvolution epifluorescence microscopy, and conjugates were imaged using DIC microscopy. Bar, 5 μm. (B) Relative retention of PKCθ-GFP constructs at the IS was quantified for 20–45 conjugates per cell line, and relative IS retention was calculated using the two ratios R1 and R2 (see Materials and Methods). *p < 0.001 in comparisons of PKCθ mutants to WT (one-way analysis of variance, Tukey's multiple comparison test).