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. 2023 Sep 15;9:95. doi: 10.1038/s41421-023-00592-6

Fig. 6. Screening of N1DARP-involving USPs-N1ICD interaction.

Fig. 6

a Total ubiquitination level of N1ICD detected by western blotting assay using Capan1 with wild-type or Y20/Y34 mutated N1DRAP. b The effect of N1DARP on the BAP1-mediated deubiquitination of N1ICD detected using western blotting assay in HEK293T. c Screening strategy to identify N1DARP-involved USPs-mediated deubiquitination of N1ICD. d Co-IP followed by western blotting assay screening N1ICD-interacting deubiquitinases in the USP family. e Co-IP followed by western blotting assay detecting polyubiquitination level of N1ICD using HEK293T transfected with N1ICD-interacting USPs selected above. f Remaining N1ICD at indicated time detected using western blotting assay in HEK293T transfected with N1ICD-interacting and deubiquitining USPs selected above after treatment with CHX. g The ubiquitination level of N1ICD detected using co-IP and western blotting assay in HEK293T transfected with N1ICD-interacting, deubiquitining, and destabilizing USPs (USP7, USP10, USP14, and USP37), followed by introduction of N1DARP. The data are representative of three independent experiments. ns no significance; *P < 0.05, **P < 0.01 by one-way ANOVA (f).