![Fig. 6. bpV[pic]-mediated activation of NFAT is dependent on TCR-proximal events. / (A) JCaM-TAg cells (p56lck-negative) were transiently transfected by the DEAE-Dextran protocol with 15 μg of the pNFAT-LUC plasmid plus 30 μg of either the control vector pEFneo (■) or the p56lck-encoding vector pEFneoLckWT (■). After a 24-hour incubation, cells were treated or not with PHA (3 μg/mL)/PMA (20 ng/mL), bpV[pic] (10 μM), or PMA (20 ng/mL)/Iono (1 μM). Cells were lysed 8 hours after stimulation and measured for luciferase activity. (B) P116 cells (ZAP-70-deficient) were transiently transfected by the DEAE-Dextran protocol with 15 μg of the pNFAT-LUC plasmid plus 15 μg of either the empty vector (■), a ZAP-70 WT (▨) or ZAP-70 kinase-dead expression vector (▪). At 24 hours after transfection, cells were treated or not with PHA (3 μg/mL), PHA (3 μg/mL)/PMA (20 ng/mL), PMA (20 ng/mL)/Iono (1 μM), or bpV[pic] (10 μM) for 8 hours and lysed. Luciferase activity was monitored as described in the “Materials and methods” section. Results are the means ± SD for triplicate samples and are representative of 2 independent experiments.](https://ash.silverchair-cdn.com/ash/content_public/journal/blood/97/8/10.1182_blood.v97.8.2390/5/h80810920006.jpeg?Expires=1722696752&Signature=O3M1jgpSEQn2fUncDvfucEzyJ1JKSeokQgNPHj9jyCW8olGY6yZVqQEJDiWUBxpaf4VvGGjDbVgDHF~GYw3KoIydyNdyv9T0zQeBumgXBgd3pBcYznzpAGR9e~~2RkLXGbxN9scel8z96nC9-FsA5bRi4fCFjhfKaoaiJxuQYzAGPQTeLyGQB2n67kc5vR1OLUuTsCD2a4orxoBCaMq3Lr6GkoRnEF1eYh7CIXqyV0UwKkfEqrXB4Ye7xIoj3QmZLdw7Ns4sTsickGLl~BYupnrwGzpYwy7vTNouS-6Q~rp6kDYoGfmM7og-MKMCjSJJ3rRaMAEVt3K9dq9NW8cFxQ__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
bpV[pic]-mediated activation of NFAT is dependent on TCR-proximal events.
(A) JCaM-TAg cells (p56lck-negative) were transiently transfected by the DEAE-Dextran protocol with 15 μg of the pNFAT-LUC plasmid plus 30 μg of either the control vector pEFneo (■) or the p56lck-encoding vector pEFneoLckWT (■). After a 24-hour incubation, cells were treated or not with PHA (3 μg/mL)/PMA (20 ng/mL), bpV[pic] (10 μM), or PMA (20 ng/mL)/Iono (1 μM). Cells were lysed 8 hours after stimulation and measured for luciferase activity. (B) P116 cells (ZAP-70-deficient) were transiently transfected by the DEAE-Dextran protocol with 15 μg of the pNFAT-LUC plasmid plus 15 μg of either the empty vector (■), a ZAP-70 WT (▨) or ZAP-70 kinase-dead expression vector (▪). At 24 hours after transfection, cells were treated or not with PHA (3 μg/mL), PHA (3 μg/mL)/PMA (20 ng/mL), PMA (20 ng/mL)/Iono (1 μM), or bpV[pic] (10 μM) for 8 hours and lysed. Luciferase activity was monitored as described in the “Materials and methods” section. Results are the means ± SD for triplicate samples and are representative of 2 independent experiments.
