Table 1.
Incubation with 75Se, hr | Total 75Se present/0.6 ml of extract
|
|||
---|---|---|---|---|
Control T cells,
|
Tat-treated T cells,
|
|||
cpm × 106 | % | cpm × 106 | % | |
28 | 0.3 | 0.5 | 1.02 | 1.73 |
50 | 1.2 | 2.05 | 2.85 | 4.85 |
Jurkat T cells were propagated in 6-ml cultures containing RPMI medium 1640 with 10% fetal bovine serum and either maltose-binding protein (42 kDa), 10 μg, (controls) or maltose-binding-Tat fusion protein (52 kDa), 10 μg, (Tat-treated cells) for 24 or 48 hr at 37°C in a CO2 incubator. Then 58.8 × 106 cpm 75SeO32− (ca. 3 nmoles; Los Alamos National Laboratory, NM) were added to each 6-ml culture and incubation was continued for 28 hr (24-hr culture) or 50 hr (48-hr culture). Cells were harvested, washed twice, and suspended in SDS/PAGE sample buffer with mercaptoethanol as described in Materials and Methods. Extracts (0.6 ml) were heated at 100°C for 10 min.