Inhibition of BMMC migration induced by SPE-7 IgE, IgE + Ag, or SCF by various pharmacologic inhibitors
Target . | Inhibitor . | SPE-7 IgE . | IgE+Ag . | SCF . |
|---|---|---|---|---|
| Src PTKs | PP2 | 2 μM | 10 μM | 0.5 μM |
| Syk | Piceatannol | 100 μM | 120 μM | > 200 μM |
| ER-27319 | 20 μM | 20 μM | > 100 μM | |
| Btk | Terreic acid | > 20 μM | > 20 μM | > 20 μM |
| PI3K | Wortmannin | 5 nM | 40 nM | 30 nM |
| LY294002 | 7 μM | 10 μM | > 50 μM | |
| MEK | PD98059 | > 50 μM | > 50 μM | > 50 μM |
| JNK | SP600125 | 30 μM | 40 μM | 40 μM |
| p38 | SB203380 | 5 μM | 2 μM | 20 μM |
| PKC | Ro31-8425 | 4 μM | 8 μM | 20 μM |
| cPKC | Gö-6976 | 2 μM | 0.5 μM | 2 μM |
| PKA | PKI | > 100 nM | > 100 nM | > 100 nM |
| KT5720 | > 100 nM | > 100 nM | > 100 nM | |
| Rho-K | Y-27632 | > 10 μM* | > 10 μM* | > 10 μM* |
| F-actin | Cytochalasin D | 0.02 μM | 0.02 μM | 0.08 μM |
| Ca2+ | EGTA | 1 mM | 1 mM | 2 mM |
| IgV | DNP-lysine | 0.4 μM | 0.4 μM | > 100 μM |
Target . | Inhibitor . | SPE-7 IgE . | IgE+Ag . | SCF . |
|---|---|---|---|---|
| Src PTKs | PP2 | 2 μM | 10 μM | 0.5 μM |
| Syk | Piceatannol | 100 μM | 120 μM | > 200 μM |
| ER-27319 | 20 μM | 20 μM | > 100 μM | |
| Btk | Terreic acid | > 20 μM | > 20 μM | > 20 μM |
| PI3K | Wortmannin | 5 nM | 40 nM | 30 nM |
| LY294002 | 7 μM | 10 μM | > 50 μM | |
| MEK | PD98059 | > 50 μM | > 50 μM | > 50 μM |
| JNK | SP600125 | 30 μM | 40 μM | 40 μM |
| p38 | SB203380 | 5 μM | 2 μM | 20 μM |
| PKC | Ro31-8425 | 4 μM | 8 μM | 20 μM |
| cPKC | Gö-6976 | 2 μM | 0.5 μM | 2 μM |
| PKA | PKI | > 100 nM | > 100 nM | > 100 nM |
| KT5720 | > 100 nM | > 100 nM | > 100 nM | |
| Rho-K | Y-27632 | > 10 μM* | > 10 μM* | > 10 μM* |
| F-actin | Cytochalasin D | 0.02 μM | 0.02 μM | 0.08 μM |
| Ca2+ | EGTA | 1 mM | 1 mM | 2 mM |
| IgV | DNP-lysine | 0.4 μM | 0.4 μM | > 100 μM |
BMMCs were preincubated with pharmacologic reagents for 30 minutes before being placed in upper wells at the time of addition of 10 μg/mL SPE-7 IgE or 100 ng/mL SCF to lower wells. For IgE+Ag stimulation, BMMCs were sensitized by overnight incubation with 0.5 μg/mL of 206 IgE. BMMCs washed with buffer were preincubated with inhibitors for 30 minutes before being placed in upper wells at the time of addition of 10 ng/mL DNP-HSA. Concentrations for 50% inhibition (IC50) are indicated.
Although BMMCs were insensitive to Y-27632, MC/9 mouse mast cells were sensitive with IC50s of 5 μM to 10 μM for SPE-7-, IgE + Ag-, and SCF-induced migration.