Fig. 4

Interaction of myosin Va (myoVa) and myosin VI (myoVI) with Na⁺/K⁺-ATPase α1 subunits endogenously expressed in HEK293 cells. HEK293 cell lysates (In, lane 1 in a and b) were precleared (PC) with rabbit IgG (a) or mouse IgG1 (b) prior to immunoprecipitation (IP, lane 3) using antibodies for myoVa (a) or myoVI (b). Loading of PC complexes in the gel preceded those of the IP complexes. Immunoprecipitation of myoVI (b (iii)), but not those of myoVa (a (iii)), led to the co-immunoprecipitation of Na⁺/K⁺ ATPase α1 subunits ((i) in a and b) expressed in HEK293 cells. Neither myoVa nor myoVI co-immunoprecipitated β-actin ((ii) in a and b) expressed in HEK293 cells. Denatured mouse IgG-HC and IgG-LC separated from their intact immunoglobulins (that is used for PC or IP) are seen in B as the blot sections were probed with mouse anti- Na⁺/K⁺ ATPase α1 and anti-β-actin antibodies