Fig. 4: USP10 promotes contractile ring localization and cytokinesis by stabilizing ANLN protein. | Cell Death & Differentiation

Fig. 4: USP10 promotes contractile ring localization and cytokinesis by stabilizing ANLN protein.

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

Fig. 4

A KYSE150 cells transfected with siRNAs were synchronized to different time periods by thymidine. ANLN and USP10 levels were detected by western blotting. Cyclin B1, E2, and phospho-histone H3 (S10) were used as markers for the different cell-cycle phases. B KYSE150 cells were subjected to DTB and released for different time periods. Interaction between ANLN and USP10 was detected by immunoprecipitation. Cyclin B1 and E2 were used as markers for the different cell-cycle phases. C The localization of ANLN and USP10 was detected by immunofluorescence in synchronized KYSE150 cells. D KYSE150 cells transfected with siRNAs were synchronized in G2/M-phase, released for 3 h, and analyzed by flow cytometry. E KYSE150 and KYSE30 cells were transfected with siRNAs for 48 h. The indicated antibodies were used for western blotting. F KYSE150 cells stably expressing HA-vector or HA-ANLN was transfected with a USP10 siRNA pool, then treated with thymidine for 24 h, and released for 12 h to obtain cytokinesis cells. Cells were fixed and detected by immunofluorescence. G Mean intensity ratio of individual cells were plotted (F-Actin fluorescence at the equatorial cortex: pole). P < 0.0001 by Student’s t-test. H KYSE150 cells stably expressing HA-vector or HA-ANLN were transfected with a USP10 siRNA pool. The indicated antibodies were used for western blotting. All data are representative of at least three independent experiments and the results were statistically analyzed using a t-test.

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