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. Author manuscript; available in PMC: 2018 Oct 1.
Published in final edited form as: Kidney Int. 2017 Oct 18;93(2):390–402. doi: 10.1016/j.kint.2017.07.027

FIGURE 1. The B1 and B2 H+-ATPase subunits and Rhcg coexist in the same protein complex.

FIGURE 1.

Co-Immunoprecipitation was performed on rat kidney medulla using crosslinking. Panel A: Co-IP was performed with anti-Rhcg (lane 2) and anti-B1 (lane 5) polyclonal antibodies or with an isotypic IgG antibody (lanes 1 and 4). Lanes 3 und 6 were loaded with 50 mg rat kidney medulla protein. Immunoblotting (IB) was performed against the B1 H+-ATPase subunit (left side) or against RhCG (right side). Panel B: the Co-IP membrane was stripped and reblotted with antibodies against Rhcg and B1 H+-ATPase antibodies as control for the IP. The upper band for B1 (approx 70 kDa) is considered unspecific. Panel C: Co-IP was performed with anti-Rhcg (lane 2) and anti-B2 (lane 5) polyclonal antibodies or with an isotypic IgG antibody (lanes 1 and 4). Lanes 3 und 6 were loaded with 50 mg rat kidney medulla protein. Immunoblotting (IB) was performed against the B2 H+-ATPase subunit (left side) or against RhCG (right side). Panel D: the Co-IP membrane was stripped and reblotted with antibodies against Rhcg and B2 H+-ATPase antibodies as control for the IP. Panel E: Coimmunostaining of Rhcg and the E H+-ATPase subunit in kidney tissue from Rhcg+/+ and Rhcg−/− mice. Immunostaining of E H+-ATPase subunit (in red) and Rhcg (green) was performed on kidney sections from Rhcg+/+ and Rhcg−/− mice. Nucleus were labelled with DAPI staining. Inserts show higher magnification of single intercalated cells.