Figure 7.
Develop inhibitory peptides against the WW-NLS in YAP1 nuclear import. (A) The structure (PDB: 2LTW) illustrates the bound state of the PPPY peptide and the first WW domain derived from YAP1. (B) The competitive binding peptide (CPY6) is developed according to the PPPY motif, and the co-IP experiment shows the interaction of the CPY6 with the wild-type or the W4A substituted WW domains of YAP1 in HEK293T cells. TCL, total cell lysates. (C) Representative images show the subcellular localization of tdWWs−3xGFP upon ectopic expression of CPY6 peptide in HEK293T cells, and the scatter chart shows their nucleocytoplasmic ratios (representative of three independent experiments, n represents the total number of the measured cells, two-tailed ANOVA test). (D) The CPY6 peptide suppresses YAP1 transcriptional activity in HEK293T cells (representative of three independent experiments, N = 3, all data shown as the average values ± SD, two-tailed Student’s t test). Representative western blotting shows YAP1 expression in luciferase reporter assays. (E) Representative images show the subcellular localization of YAP1-GFP upon ectopic expression of CPY6 peptide in HEK293T cells, and the scatter chart shows their nucleocytoplasmic ratios (representative of three independent experiments, n represents the total number of the measured cells, two-tailed ANOVA test). (F) Representative immunostaining images show the subcellular localization of endogenous YAP1 proteins upon LMB treatment (0.5 μM) in HEK293T cells following 1-wk culture with 1 μM of synthesized PPPY peptide. (G) The synthesized PPPY peptide blocks the interaction of Fc−YAP1 and His-KPNA2 in vitro. In the absence or presence of PPPY peptides (30 μM), the recombinant His-KPNA2 (3.3 μM) is incubated with Fc- or Fc-YAP1-engaged protein A/G plus agarose at room temperature for 1 h, then the pulled-down proteins by protein A/G plus agarose are analyzed by SDS PAGE. Source data are available for this figure: SourceData F7.

Develop inhibitory peptides against the WW-NLS in YAP1 nuclear import. (A) The structure (PDB: 2LTW) illustrates the bound state of the PPPY peptide and the first WW domain derived from YAP1. (B) The competitive binding peptide (CPY6) is developed according to the PPPY motif, and the co-IP experiment shows the interaction of the CPY6 with the wild-type or the W4A substituted WW domains of YAP1 in HEK293T cells. TCL, total cell lysates. (C) Representative images show the subcellular localization of tdWWs−3xGFP upon ectopic expression of CPY6 peptide in HEK293T cells, and the scatter chart shows their nucleocytoplasmic ratios (representative of three independent experiments, n represents the total number of the measured cells, two-tailed ANOVA test). (D) The CPY6 peptide suppresses YAP1 transcriptional activity in HEK293T cells (representative of three independent experiments, N = 3, all data shown as the average values ± SD, two-tailed Student’s t test). Representative western blotting shows YAP1 expression in luciferase reporter assays. (E) Representative images show the subcellular localization of YAP1-GFP upon ectopic expression of CPY6 peptide in HEK293T cells, and the scatter chart shows their nucleocytoplasmic ratios (representative of three independent experiments, n represents the total number of the measured cells, two-tailed ANOVA test). (F) Representative immunostaining images show the subcellular localization of endogenous YAP1 proteins upon LMB treatment (0.5 μM) in HEK293T cells following 1-wk culture with 1 μM of synthesized PPPY peptide. (G) The synthesized PPPY peptide blocks the interaction of Fc−YAP1 and His-KPNA2 in vitro. In the absence or presence of PPPY peptides (30 μM), the recombinant His-KPNA2 (3.3 μM) is incubated with Fc- or Fc-YAP1-engaged protein A/G plus agarose at room temperature for 1 h, then the pulled-down proteins by protein A/G plus agarose are analyzed by SDS PAGE. Source data are available for this figure: SourceData F7.

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal