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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. 4. SAMe and MTA treatment reduced Ubc9’s state of phosphorylation and sumoylation.

Fig. 4

(A) Huh-7, RKO and MCF-7 cells were treated with SAMe (2mM) or MTA (1mM) for 24 hours. Total protein phosphorylation was examined by Western blotting using anti-phospho antibody, while phospho-Ubc9 was determined by first immunoprecipitating total Ubc9, followed by Western blotting with anti-phospho antibody. Sumoylation was measured using anti-SUMO-1 antibody in Western blotting. The values are expressed as % of control cells (mean ± SEM) from 3 independent experiments. *P<0.02, **p<0.05 vs. control cells. (B) RKO cells were cultured and treated with phosphatase inhibitor cocktail (1:10000) for 24 hours and processed for immunoprecipitation using anti-Ubc9 antibody followed by Western blotting with anti-phospho antibody or anti-Ubc9 antibody. Results are expressed as % of control (mean ± SEM) from 3 independent experiments. *P<0.02 vs. control cells. (C) Prediction of potential kinases that can phosphorylate Ubc9 protein sequence using the KinasePhos, PostMod, MotifScan, Phosphonet analysis tools.