Fig. 4. Coimmunoprecipitation of TRPV channel subunits. (A-E) Plasmids encoding FLAG-tagged or YFP-tagged TRPV channels were co-transfected in HEK293 cells in the combinations indicated below the panels. One day after transfection, membranes were solubilized and lysates were immunoprecipitated (IP) with anti-FLAG antibodies. Membrane lysates (upper panels) or immunoprecipitates (middle and lower panels) were separated by SDS-PAGE. Upper and middle panels, TRPV in the membrane lysates and coprecipitated TRPV channels were detected by immunoblotting (IB) with anti-GFP antibodies. The recovery of the respective immunoprecipitated TRPV-FLAG channel is shown in the lower panels by probing blots with anti-FLAG antibodies. Arrowheads indicate the expected sizes of the respective TRPV-FLAG channel subunits.