CaMKKα enhances CKLiK activity.

(A) HA_CKLiK (8 μg) was transfected in COS cells with or without CaMKKα_VSV (2 μg). Before harvesting, cells were stimulated with or without 1 μmol/L ionomycin as indicated. HA_CKLiK was immunoprecipitated and kinase assays were performed using CREM as substrate in the presence or absence of Ca⁺⁺/calmodulin. Equal protein expression was demonstrated by Western blotting. For CKLiK antibody 12CA5 (middle panel) and for CaMKKα antibody anti-VSV (bottom panel) is used. (B) CREB_GAL4 and CKLiK (4 μg) were cotransfected without or with CaMKKα (2 μg) as indicated. CaMKKα was transfected without CKLiK as a negative control. Eighteen hours before harvesting, cells were stimulated with or without 1μmol/L ionomycin and reporter assays were performed. Data represent at least 3 independent experiments ± SEM and are corrected for transfection efficiency.