Fig. 3

Interaction of non-muscle myosin heavy chains 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 mouse IgG2b (PC, lane 2 in a and b) prior to immunoprecipitation (IP, lane 3) using antibodies for myh9 (a) and myh10 (b). Loading of PC complexes in the gel preceded those of the IP complexes. Immunoprecipitation of myh9 (a) and myh10 (b) led to the co-immunoprecipitation of Na⁺/K⁺ ATPase α1 subunits ((i) in a and b) and β-actin ((ii) in a and b) expressed in HEK293 cells. Na⁺/K⁺-ATPase α1 and β-actin immunoreactive signals in the depleted supernatant lane (DS, lane 4 in a) indicates that the ATPase survives the IP procedure. Denatured mouse IgG-HC and IgG-LC separated from their intact immunoglobulins (that is used for PC or IP) are seen in (a and b) as those blot sections were probed with mouse anti-β-actin antibodies