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HTRF cAMP assays (Cisbio) were performed following the manufacturer's instructions. For serum shift assays, cells were plated overnight in 384-well plates. On the day of the assay, growth medium was aspirated from the plates and replaced with fetal bovine serum (10 μL). Test compounds were serially diluted in DMSO and then diluted in assay buffer (PBS with 200 μM IBMX & 0.1% BSA) to achieve a 2× concentration containing 1% DMSO prior to addition to assay plates (10 μL).
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HTRF cAMP assays (Cisbio) were performed following the manufacturer's instructions. For serum shift assays, cells were plated overnight in 384-well plates. On the day of the assay, growth medium was aspirated from the plates and replaced with fetal bovine serum (10 μL). Test compounds were serially diluted in DMSO and then diluted in assay buffer (PBS with 200 μM IBMX & 0.1% BSA) to achieve a 2× concentration containing 1% DMSO prior to addition to assay plates (10 μL).
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