Figure 5.

Tpl-2 activates NFATp. (A) Relative CAT activity in freeze–thaw lysates of NIH 3T3 cells stably transfected with an SRα-based Tpl-2 construct or with SRα and cotransfected with expression constructs of NFATp, NFATc, or NFATx and the reporter construct NFAT/CAT. All transfections were performed in triplicate and transfection efficiency was monitored by using βgal. (B–D) Subcellular localization of NFATp, NFATc, and NFATx expressed from transiently transfected expression constructs in the NIH 3T3 shown in A. Localization was determined by immunofluorescence using anti-HA (HA-NFATc) or anti-Flag (Flag-NFATp or Flag-NFATx) mouse monoclonal antibodies and an anti-mouse IgG secondary antibody conjugated with fluorescein isothiocyanate. Stained cells were examined by confocal microscopy. (Left) Entire cell. (Right) Confocal image of a slice through the nucleus.