Table 1.
Pharmacologic agents influence Btk protein level
| Experiment | Btk | Treatment | Anti-IgM | Fluorescence* |
|---|---|---|---|---|
| 1 | WT | None | (−) | 4 |
| WT | None | (+) | 42 | |
| WT | LY294002 (−0.5 hour) | (+) | 8 | |
| WT | LY294002 (+1 hour) | (+) | 8 | |
| WT | LY294002 (+3 hours) | (+) | 36 | |
| 2a | WT | None | (−) | 9 |
| WT | None | (+) | 28 | |
| WT | PMA + ionomycin | (−) | 28 | |
| 2b | xid | None | (−) | 5 |
| xid | None | (+) | 7 | |
| xid | PMA + ionomycin | (−) | 18 |
Influence of activators or inhibitors of intracellular signaling pathways on Btk protein expression. Experiment 1: Balb/c splenic lymphocytes were stimulated by cross-linking BCR with anti-IgM (20 μg/ml) for 4 hours at 37°C. LY294002 (20 μM) or carrier (DMSO 1%) alone was added 0.5 hours before, 1 hour after, or 3 hours after receptor crosslinking. Experiment 2: Balb/c (a) or Balb/xid (b) splenic lymphocytes were stimulated by cross-linking BCR with anti-IgM (20 μg/ml), or by treatment with phorbol myristic acid (10 nM) and ionomycin (100 nM) or carrier (DMSO 1%) alone, and then were incubated for 4 hours at 37°C. Cells were fixed and immunostained with a nonspecific antibody or anti-Btk followed by PE-anti-rabbit. The mean channel fluorescence intensity for each sample was rounded to the nearest whole number. PMA, phorbol myristic acid; WT, wild type.
*Mean channel fluorescence intensity.