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Fig. 6. NF-AT-dependent transcription in LAT-deficient cells reconstituted with different mutants. The NF-AT-Luc plasmid and the indicated LAT constructs were transfected into JCAM2.5 cells. 18 hours after transfection, transfected cells were incubated in standard round-bottom 96-well plates without or with Raji B cells (ratio 0.5 B cell for 1 T cell), either unpulsed or pulsed with 1 µg/ml of SEE. Luciferase activity was assayed 6 hours later using a Promega luciferase assay kit according to the manufacturer's instructions. The percentages and mean fluorescence of GFP-transfected cells were checked by FACS analysis to ensure similar transfection efficiencies. One representative experiment out of three is presented.
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