FIGURE

Fig. 10

ID

ZDB-FIG-240905-10

Publication

Chen et al., 2024 - NEDD8 enhances Hippo signaling by mediating YAP1 neddylation

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Fig. 10

Xiap catalyzes Yap1 neddylation at lysine 159 (K159). A, neddylation of yap1 is detected by immunoprecipitation with Ni-NTA agarose and Western blot in HEK293T cells transfected with His-nedd8 and WT Myc-yap1 or one of the site-mutated yap1 constructs for 24 h. B, protein level of yap1 in nuclei is detected by Western blot analysis in HEK293T cells transfected with Myc-yap1-WT, K62R, K159R, or K208R/K210R. ?-Tubulin and H3 are used as markers for the cytoplasmic and nuclear fractions, respectively. C, fluorescence of Myc is detected by confocal microscopy in HEK293T cells transfected with Myc-yap1 or yap1 site-mutant plasmids (K62R, K159R, K208R/K210R) for 24 h. The scale bar represents 100 ?m. D, relative luciferase activity of the 8xGTIIC-Luc promoter in HEK293T cells expressing the indicated constructs together with or without His-nedd8. Four separate experiments were performed, with each measurement performed at least three times. E, neddylation of yap1 is detected by immunoprecipitation with Ni-NTA agarose and Western blot in HEK293T cells transfected with Myc-yap1-WT or Myc-yap1-K159R for 24 h. Data are expressed as mean � SD. ?p < 0.05, ??p < 0.01, and ???p < 0.001.IB, immunoblotting; IP, immunoprecipitation; TCL, total cell lysate; ns, not significant; YAP1, yes-associated protein 1.

Expression Data

Expression Detail

Antibody Labeling

Phenotype Data

Phenotype Detail

Acknowledgments

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