Figure 3.

Coimmunoprecipitation of Cdc50p/Lem3p family members with Drs2p/Neo1p family members. Cells were grown at 25°C to a cell density of 0.5 OD₆₀₀/ml in YPDA medium. Membrane extracts were then prepared as described in the MATERIALS AND METHODS. Myc-tagged P-type ATPases were immunoprecipitated with an anti-Myc antibody from membrane extracts. Immunoprecipitates were subjected to SDS-PAGE, followed by immunoblot analysis using antibodies against HA (top), Myc (middle), and Lem3p (bottom). Arrows indicate the bands of anti-Myc antibody polypeptides detected by secondary antibodies (anti-mouse IgG-HRP). The results shown are representatives of several experiments. The strains used were as follows: YKT755 (Drs2p-Myc Cdc50p-HA) and YKT283 (Cdc50p-HA) (A), YKT759 (Dnf1p-Myc Cdc50p-HA) and YKT283 (Cdc50p-HA) (B), and YKT761 (Neo1p-Myc Cdc50p-HA) and YKT283 (Cdc50p-HA) (C).