Fig. 7.

NFAT1 protein stability is not affected by IRF-2BP2. (A) HEK293T cells were transfected with pLIRES-EGFP-CA-NFAT1 (2 μg) and increasing amounts of pcDNA4-IRF-2BP2 (0.25 to 2 μg). After 48 h, the indicated proteins were detected by Western blotting with specific antibodies. Results are representative of three independent experiments. (B) Jurkat cells were electroporated with the luciferase reporter vector 3× NFAT-Luc (1 μg), empty vectors (8 μg), the pcDNA5-NFAT1 vector (8 μg), and increasing amounts of the pcDNA4-IRF-2BP2 vector (1 to 8 μg). After 24 h, cells were left untreated or were treated with MG132 (20 μM) and stimulated for 6 h with PMA (10 nM) plus ionomycin (1 μM). The firefly luciferase reporter gene was normalized with a renilla vector (0.1 μg pRL-TK). Results represent means from three independent experiments ± standard deviations.