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. 2017 Feb 14;8(11):18248–18259. doi: 10.18632/oncotarget.15319

Figure 2. NICD induce ubiquitination and degradation of ATXN1.

Figure 2

A. Upper panel: Western blotting analysis of HeLa cells transfected with Myc-NICD and treated with 100 μg/ml cycloheximide for various times. Lower panel: Density of ATXN1 was measured and plotted. The half-life of ATXN1 is indicated by dot lines. B. Western blotting analysis of HEK293 cells transfected with HA-ATXN1, Myc-NICD, or both, followed by treatment with MG132 for 6 h. C. Lysates prepared from HEK293 cells co-transfected with HA-ATXN1 and Myc-NICD were immunoprecipitated using an anti-Myc antibody, and the resulting immunoprecipitates were subjected to western blotting analysis. D. Upper panel: ATXN1 deletion constructs. Lower panel: HEK293 cells were co-transfected with Xpress-ATXN1 deletion mutants and Myc-NICD. After transfection, cell lysates were immunoprecipitated with an anti-Myc antibody and subjected to western blotting analysis. E. HEK293 and HeLa cells were co-transfected with Myc-NICD and HA-ATXN1 with or without Xpress-Ub and treated with MG132 for 6 h. ATXN1 was immunoprecipitated using an anti-HA antibody. Cell lysates were analyzed via western blotting with the indicated antibodies. IP (Immunoprecipitation), IB (Immunoblot), XP (Xpress), HC (Heavy chain).