Determination of the optimal conditions for the screening of active PIs.
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Clonal SupT1 cells harboring inducible Gal4, PRm/Gal4, or PR/Gal4 were activated in a 96 well-plate format under various conditions. (A) Doxycycline titration. Cells were pre-incubated with either DMSO or 10µM IDV and then either left untreated, or activated with 50, 100, 200, 500, 1,000, or 2,000ng/mL of Dox. Cells were analyzed by flow cytometry to determine the number of eGFP positive cells. (B) Time course analysis of eGFP induction in response to Indinavir with or without Dox. Cells were pre-incubated with 10µM IDV and then either left untreated, or activated with 1µg/mL Dox. Cells were analyzed 4, 8, 12, 16, 20, 25, 50 or 75 hours after activation by flow cytometry.