Fig. 3: USP10 removes K11- and K63-linked ubiquitin chains of ANLN through its deubiquitinase activity. | Cell Death & Differentiation

Fig. 3: USP10 removes K11- and K63-linked ubiquitin chains of ANLN through its deubiquitinase activity.

From: Targeting USP10 induces degradation of oncogenic ANLN in esophageal squamous cell carcinoma

Fig. 3

A KYSE150 cells were transfected with siRNAs for 24 h and then treated with cycloheximide (CHX) (10 μg/ml) for different times. ANLN protein level was detected by western blotting. B KYSE150 cells stably expressing HA-vector or HA-USP10 were treated with CHX (10 μg/ml) for different times. ANLN protein level was detected by western blotting. C KYSE150 cells transfected with siRNAs were treated with MG132 (20 μM) for 10 h before harvesting, and ANLN protein levels were detected by western blotting. D KYSE150 cells transfected with siRNAs and HA-Ub were treated with MG132 (20 μM) for 10 h before harvesting, and the ubiquitination level of endogenous ANLN was measured using an ubiquitination assay. E HEK293T cells were co-transfected with Flag-ANLN, HA-Ub, USP10-WT, or CA, and treated with MG132 (20 μM) for 8 h before harvesting. Ubiquitination assays were used to detect the ubiquitination level of Flag-ANLN. F HEK293T cells were transfected with the indicated plasmids and treated with MG132 (20 μM) for 8 h before harvesting. An ubiquitination assay was used to detect the ubiquitination level of Flag-ANLN. All data are representative of at least three independent experiments.

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