WWL70

One hundred micrograms per milliliter of BV2 microsomes are pre-incubated with WWL70 for 5 min at 23°C, then mixed with 10 μM of Arachidonic acid (AA) for 1 min at 23°C. 500 μg/mL brain microsomes are incubated with 10 μM of AA for 2 min at 23°C. The reaction is stopped by mixing with stannous acid (5 mg/mL in 0.1 N HCl) to deactivate the enzyme and convert intermediate PGH₂ to PGF_2α, followed by the measurement of PGE₂ concentration by Enzyme-linked immunoassay (EIA). The activity is determined after subtraction with the amount of PGE₂ in the microsome fraction incubated without substrate^[2].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

BV2 cells (90% confluence) in 10-cm dishes are treated with WWL70 (10 μM) for 1 h. After rinsing with PBS once, the cells are collected by centrifugation at 5000 g for 2 min. The pellet is suspended with 0.1 mL of 0.02% trifluoroacetic acid (TFA) and 1 nmol of 2-AG-d₈ by pipetting and dispersed in 4 mL of acetonitrile in a silanized glass tube to precipitate the debris overnight at -20°C. The supernatant after centrifuged at 5000 g for 5 min is transferred to a new glass tube and evaporated under a nitrogen gas stream in a mild hot water bath (approximately 35°C). 2-AG is resuspended with 0.1 mL of acetonitrile and stored at -80°C until mass analysis^[2].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Seven-week-old, male C57BL/6 mice weighing 25 to 30 g are used in this study. Animals are maintained under a controlled environment with a temperature of 23±2°C, a 12 h light/dark cycle, and access to food and water ad libitum. WWL70 (5 mg/kg or 10 mg/kg) in physiologic saline or an equal volume of 1% DMSO in saline (10 mL/kg) is injected intraperitoneally, and then once a day for 3, 7, or 21 days depending on the experimental design. During the 21-day treatment regimen, animals are subjected to a battery of behavioral tests at different time points. Two hours after the last injection on day 21 post-injury, animals are sacrificed and brain tissues are collected for histological analysis^[3].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.