Fig. 6

Recombinant myh9 co-immunoprecipitate Na⁺/K⁺-ATPase α1 subunits expressed in HEK293 cells. Lysates of HEK293 cells transiently transfected with GFP (In, lane 1) or myh9 tagged with GFP in the C-terminus (In, lane 4; myh9-GFP) were precleared (PC) with mouse IgG1 antibodies (a, lanes 2 and 5) or goat immunoglobulins (gIgG) (b, lanes 2 and 5) prior to immunoprecipitation (IP) using mouse anti-GFP antibodies of the IgG1 isotypes (a, lanes 3 and 6) or goat anti-GFP antibodies (b, lanes 3 and 6). Loading of PC complexes in the gel preceded those of the IP complexes. Na⁺/K⁺-ATPase α1 immunoreactive bands in lanes 1, 4 and 6 but not in any other lanes ((i) and (iii)) indicated co-immunoprecipitation of Na⁺/K⁺-ATPase α1 subunits from HEK293 cells transfected with myh9-GFP plasmids but not from those transfected with GFP plasmids. As expected immunoprecipitation using anti-GFP antibodies led to the co-immunoprecipitation of β-actin (lane 6 in (ii) and (iv)) from HEK293 cells transfected with myh9-GFP plasmids but not from those transfected with GFP plasmids. Immunoprecipitation using goat anti-GFP antibodies (b) led to a cleaner co-IP of Na⁺/K⁺-ATPase α1 subunits from HEK293 cells expressing myh9-GFP. Full length images of western blots are presented in Figure Additional file 4: Figure S3B (for part a) and Additional file 5: Figure S4B (for part b)