Fig. 6.
Hepatic Cyp2a5 UPD and ALD: a major role of gp78 in Cyp2a5 ubiquitination. (A) Cycloheximide-chase analyses of hepatocytes from three individual WT and gp78-KO mice were cultured, PB-pretreated for 3 days, and on day 4 treated with or without the proteasomal inhibitor bortezomib (BTZ) or the ALD-inhibitors 3-MA/NH4Cl for 0, 8, and 24 hours before cell harvesting. Cell lysates were immunoblotted for Cyp2a or gp78 with actin as a loading control. Densitometric quantification (mean ± S.D.) of individual cultures from three mice. Statistical significance determined by the Student’s t test, ***P < 0.001 vs. WT at 0 hour; ##, ###P < 0.01, 0.001 vs. WT+ CHX; §P < 0.05 vs. KO at 0 hour. (B) Representative Ub-immunoblotting analyses of Cyp2a-immunoprecipitates from aliquots of PB-pretreated cultured cell lysates obtained from two of the three different mice treated with or without BTZ as in (A).