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Superose 6 (S6) gel-filtration chromatography of HEK cell extracts. HEK293 cells transfected with FLAG-tagged PDE3A1 or PDE3A2 were treated with 1 µM isoproterenol for 90 s or with 10 ng/mL PMA for 15 min, respectively. Solubilized cell extracts (3 mg protein, 1 mL) were subjected to chromatography on S6 columns. Then 10-μL portions of eluted fractions (0.5 mL) were assayed for PDE3 activity, and 20-μL portions were used for SDS/PAGE and Western blot analysis with anti-PDE3A antibodies.
