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. 2010 Mar 7;87(3):440–448. doi: 10.1093/cvr/cvq077

Figure 2.

Figure 2

PMCA interacts with eNOS through the proximal part of the catalytic domain. Mapping of the region of PMCA involved in the interaction with eNOS. Expression vectors encoding Flag-tagged PMCA2b(462–684), PMCA2b(1143–1243), PMCA4b(1–92), PMCA4b(428–651), or PMCA4b(652–840) were transfected in HEK 293 cells. Transfection of p3xFlag-CMV7.1 empty vector (Sigma) (F-empty) was used as a negative control. Protein lysates were incubated with commercial recombinant eNOS (Sigma) (10 μM, final concentration), and complexes were precipitated with an anti-eNOS polyclonal antibody (Sigma). Flag-tagged immunoprecipitated proteins (upper panel) and the expression levels of the Flag-tagged proteins prior to immunoprecipitation (lower panel) were analysed by western blot with anti-Flag antibody.