Figure 7. Src kinase does not coimmunoprecipitate with α-subunit in MCF10A or MDA-MB-231 cells.

MCF10A and MDA-MB-231 cells were treated with (+) or without (-) 500 nM ouabain for 24 hours. Lysates were collected and subject to immunoprecipitation using a rabbit anti-α antibody, raised against Na,K-ATPase α-subunit’s M4/M5 cytoplasmic loop. 10% of the initial lysate (IN), the co-immunoprecipitated proteins (IP), and 10% of the unbound supernatant (Sup) after IP were used for western blots. Western blot analysis was performed and probed using mouse anti-Na,K-ATPase α₁-subunit, mouse anti-Src, and mouse anti-Na,K-ATPase β₁ subunit antibodies.