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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. 7. Mutation at Ubc9 S71 reduced interaction of Cdc2 with Ubc9 and Ubc9 protein stability.

Fig. 7

(A) RKO cells were transfected with the vector, hUbc9-WT, hUbc9-F70mut, and hUbc9-F71mut for 48 hours. Co-IP analysis was done as described in Methods for phosphorylated, total Ubc9 and Cdc2. Results are expressed as % of WT (mean ± SEM) from 4 independent experiments. *P<0.02 vs. WT. (B) Ubc9 protein stability was determined in RKO and Huh-7 cells by Western blot analyses of HA tag following cycloheximide (CHX) treatment as described in Methods. Representative blots are shown. (C) Protein stability was determined using linear regression and half-life calculated using equation indicated. Results represent mean±SEM from 3 independent experiments expressed as % of respective 0 hr level for both cell types (p<0.05 between WT and MUT).