Figure 4.
SPAK neither activates nor binds to AE2 (A and B) HeLa cells were transfected with AE2 only or AE2 + SPAK. CBE activity was assessed by the slope of pHi in the absence of Cl− at the beginning of time course (30 ˜45 sec) and height to reach the maximum pHi point from the minimum point. The bars show the mean ± SEM. (D) The Co-IP of SPAK with AE2. NBCe1-B was tagged with GFP and SPAK was tagged with flag. Anti-GFP, anti-HA, and anti-flag antibodies were used for the Co-IP. Input β-actin was used as a loading control. The (-) represented cell lysates.