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. Author manuscript; available in PMC: 2013 Sep 1.
Published in final edited form as: Hepatology. 2012 Jul 12;56(3):982–993. doi: 10.1002/hep.25701

Fig. 5. Cdc2 phosphorylates Ubc9 in vitro, interacts with Ubc9 in vivo and its expression is inhibited by SAMe and MTA.

Fig. 5

(A) Cdc2/Cyclin B phosphorylates recombinant Ubc9 in vitro. Results are expressed as % control (mean ± SEM) from 3 independent experiments. *P<0.001 vs. Ubc9, †p<0.05 vs. Ubc9+Cdc2/CyclB. (B) RKO cells were treated with SAMe and MTA for 24 hours. Ubc9 expression was examined by real-time PCR and Western blotting. Results are expressed as fold or % control (mean ± SEM) from 3 independent experiments. *P<0.001 vs. control cells. (C) Effect of Cdc2 RNAi, SAMe and MTA treatments on Ubc9 and Cdc2 protein levels. RKO cells were treated with scrambled or Cdc2 RNAi or with 2mM SAMe or 1mM MTA as described in Methods. Results are expressed as % of scrambled control (mean ± SEM) from 3 independent experiments. *P<0.01, **p<0.001 vs. scrambled control. (D) Ubc9 interacts with Cdc2 in vivo. Immunoprecipitation with anti-Ubc9 antibody followed by Western blotting for phospho-Ubc9, total Ubc9, and Cdc2 in Mat1a KO livers was done as described in Methods. Results are expressed as % of WT mice (mean ± SEM) from 4 mice per group. *P < 0.001 vs. WT; **P < 0.04 vs. KO.