Fig. 4.
Increased CLP-5LO coimmunoprecipitation in ionophore-stimulated MM6 cells. Differentiated control and FLAP knockdown MM6 cells (∼2 × 106 cells) in PGC buffer were stimulated with ionophore A23187 (5 µM) as indicated for 5 min and then lysed (Materials and Methods). Equal amounts of protein lysates were subjected to immunoprecipitation with either normal rabbit IgG or anti-5LO antibody. 5LO and CLP in the lysates (inputs) and in immunoprecipitates were detected by immunoblot analysis with anti-5LO and anti-CLP antibodies. Similar results were obtained for control MM6 cells in four additional independent experiments and for FLAP knockdown cells in one additional experiment.