TABLE 2.

Pharmacological inhibition of endozepine production

Astrocytes in HSCC were preincubated for 1 h with the indicated inhibitors prior to induction by either 50 mm KCl, 100 nm cortisol or 2 μg/μl of rapamycin. The amount of endozepine produced was quantified from 400 μl of buffer harvested after induction for 15 (potassium chloride, cortisol) or 30 min (rapamycin). The following concentration of inhibitors were used: 10 μg/ml of brefeldin A, 200 ng/ml of pertussis toxin, 20 μm mPKI, 100 nm Akt inhibitor IV, 10 μm PD59059, 1 μm U73122, 1 μm Mifepristone, 2 mm 3-methyladenine. Each experiment was repeated at least three times.

	Potassium chloride	Cortisol	Rapamycin
Brefeldin A	No inhibitiona	No inhibition	No inhibition
Pertussis toxin	No inhibition	No inhibition	No inhibition
mPKI	100% inhibition	100% Inhibition	100% Inhibition
Akt inhibitor IV	No inhibition	No inhibition	100% Inhibition
PD 98059	No inhibition	No inhibition	90% Inhibition
U72122	No inhibition	No inhibition	No inhibition
Mifepristone	No inhibition	98% Inhibition	Not done
3-Methyladenine	Not done	95% Inhibition	95–98% Inhibition

^a No inhibition indicates that the endozepine detected was indistinguishable from maximal level of 80 pmol/mg of protein, whereas 100% inhibition indicates the activity was under the detection limit of 0.5 pmol of endozepine/mg of protein.