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. Author manuscript; available in PMC: 2014 Dec 23.
Published in final edited form as: Nat Commun. 2014 Jun 23;5:4234. doi: 10.1038/ncomms5234

Figure 6. CDK6 sumoylation inhibits its ubiquitination and degradation.

Figure 6

(a) LN229 cells, transduced or not with the indicated shRNAs, were treated with MG132 for the indicated hours (top) and examined by western blotting using the indicated antibodies (left). (b) The transduced cells were immunoprecipitated (IP) by a CDK6 antibody. The IP and whole cell lysate (WCL) were examined by western blotting using UB and another CDK6 antibody. (c) The half-life of Flag-CDK6 protein was evaluated by western blotting (top) and quantified against the control β-actin protein in SUMO1 and control shRNA expressing cells (bottom). (d) SUMO1 shRNA expressing LN229 cells were transfected with Flag-CDK6 wt and K-R mt with HA-K48-polyubiquitin (HA-K48-UB). Flag immunoprecipitates were examined by western blotting using HA and CDK6 antibodies. (e) The SUMO1-613 shRNA expressing LN229 cells were transfected with Flag-CDK6 wt, K147R and empty vector and analyzed by western blotting (top) and cell growth assay (bottom) (points: means; bar: SD; n = 6 from two independent experiments; *** P < 0.001; student’s t-test).