Fig. 5. SDF2 is an Hsp90 client protein and interacts with its M domain.
(A and B) Representative immunoblot analyses (left) and averaged densitometric data (right) show total Akt, eNOS, SDF2, and Hsp90 abundance in HUVECs (A) and COS7 cells (B) treated with GA or vehicle [dimethyl sulfoxide (DMSO)]. (C) Representative immunoblot analyses show the expression of HA, eNOS, and Hsp90 in whole-cell lysates (WCL) from COS7 cells cotransfected with SDF2-HA and eNOS or empty vector. HA immunoprecipitates were blotted for eNOS and Hsp90. (D) Representative immunoblot analyses show the expression of FLAG, eNOS, and HA in WCL from COS7 cells cotransfected with SDF2-HA, eNOS, or empty vector and FLAG-tagged deletion mutants of Hsp90β (as indicated). FLAG immunoprecipitates (IP: FLAG) were blotted for FLAG, eNOS, and HA. Actin was used as a loading control in (A) to (D). Data are means ± SEM. n = 3 independent experiments for (A) to (D). *P < 0.05 compared to DMSO.