Figure 3. Mutation in the PDZ-binding motif of hSlo-β1 prevented hSlo-β1 ubiquitination and degradation.

Sequence alignment of human, rabbit, cattle, rat, and mouse KCNMB1 cDNA shows a conserved PDZ-binding motif (A). ▼: applied mutation site. 72-h after co-transfection with atrogin-1 and ubiquitin, Flag-hSlo-β1V146A expression was significantly higher than that of Flag-hSlo-β1 wt (B). Immunoprecipitates of anti-Flag antibody against HEK293 cell lysates with following transfection conditions: no transfection, Flag-hSlo-β1/ubiquitin, Flag-hSlo-β1/atrogin-1/ubiquitin, Flag-hSlo-β1V146A/atrogin-1/ubiquitin and Flag-hSlo-β1/atrogin-1ΔF/ubiquitin, were resolved and blotted against anti-ubiquitin antibody (C).