Additional file 8: of Multiple myosin motors interact with sodium/potassium-ATPase alpha 1 subunits

Figure S7. Lack of co-immunoprecipitation of Na+/K+-ATPase α1 subunits and β-actin by recombinant myoVa. Lysates of HEK293 cells transiently transfected with GFP or myoVa-GFP plasmids were precleared with rabbit IgG (PC; lanes 2 and 5 in A) or goat IgG (PC; lanes 2 and 5 in B) prior to immunoprecipitation using rabbit anti-GFP antibodies (IP; lanes 3 and 6 in A) or goat rabbit anti-GFP antibodies (IP; lanes 3 and 6 in B). Loading of PC complexes in the gel preceded those of the IP complexes. Presence of Na+/K+-ATPase α1 immunoreactive bands in lanes 1 and 4, and absence of any Na+/K+-ATPase α1 immunoreactive bands in lanes 2, 3, 5 and 6 (panel (i) in A and B) indicated lack of co-immunoprecipitation of Na+/K+-ATPase α1 subunits from HEK293 cells transfected with myoVa-GFP. MyoVa-GFP did not co-immunoprecipitate β-actin (lane 6 in (ii) in A and B) from HEK293 cells. Staining the blots with mouse anti-GFP antibodies (NeuroMab: 75–131) indicated successful immunoprecipitation of GFP (lane 3 in (iii) in A and B) and myoVa-GFP (lane 6 in (iv) in B) from HEK293 cell lysates. (TIF 1906 kb)