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HEK293T cells were seeded on 12-well plates, allowed to recover overnight and treated for the indicated times with the indicated compounds.
The indicated compounds in different concentrations (100 μl) were added to cells.
IC50 values indicated compounds 1 3 exhibited moderate or less cytotoxic activity in vitro.
For treatment experiments, NSCs were treated with 0.1% DMSO, indicated compounds (10 μM), and puromycin (positive control) (10 μM).
Kinetic and molecular modeling studies also indicated compounds were mixed-type inhibitors, binding simultaneously to active, peripheral and mid-gorge sites of AChE.
Radioligand binding assays indicated compounds 6f, 6b, and 9f display potent in vitro binding affinities with nanomolar Ki values and at least 100 2000-fold selectivity for CB2.
The ratio of the wavelengths (F340/F380) reflects changes in the internal calcium concentration after perfusion with the indicated compounds.
Cells were allowed to migrate for 6 h in a humidified chamber at 37°C with 5% CO2 in the absence or presence of the indicated compounds.
For a Ras activation-redistribution assay, COS-7 cells cultured on glass bottom dishes from Mattek Corp. (Ashland, MA) were transiently transfected with YFP-RBD and serum starved for 16 h, after which they were stimulated with the indicated compounds.
Plates were supplemented with the indicated compounds.
Where indicated, compounds or proteins were added to the reactions.
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