Journal: bioRxiv

Article Title: An improved TEAD dominant-negative protein inhibitor to study Hippo YAP1/TAZ-dependent transcription

doi: 10.1101/2024.10.03.615022

Figure Lengend Snippet: A . Model showing Hippo signaling and how TEADi disrupts YAP1/TAZ binding to TEAD transcription factors, specifically in the nucleus. The building blocks of TEADi are shown. TBD= TEAD-binding domain, NLS= nuclear localization signal. B . Detail of the TBDs that compose the TEAD inhibitor. Colors indicate the stretch of the amino acid sequence from each protein found in TEADi. Reported post-transcriptional modifications in TBDs are indicated ( www.phosphosite.org ). p= phosphorylation, ub= ubiquitination, sm= SUMOylation, ac=acetylation. The homologous site in YAP1 and TAZ that binds TEAD is underlined. C . Detail of the homologous site in YAP1 and TAZ that binds TEAD and mutations introduced in TEADiv1 and TEADiv2. D . Depiction of the building blocks for constructions used in this study (not at scale).

Article Snippet: Protein visualization was performed using the following antibodies: anti-GAPDH (Cell Signaling; clone no. 14C10; catalogue no. 2118; 1:2000), anti-GFP (Cell Signaling; clone no. D5.1; catalog no. 2956; 1:2000), anti-HA tag antibody (Cell Signaling; clone no. C29F4; catalog no. 3724; 1:1000), YAP1 (Cell Signaling; clone no. D8H1X; catalog no. 14074; 1:1000), TAZ (Cell Signaling; clone no. V386; catalog no. 4883; 1:1000), CYR61 (Cell Signaling; clone no. D4H5D; catalog no. 14479; 1:1000), and Pan-TEAD (Cell Signaling; clone no. D3F7L; catalog no. 13295; 1:1000).

Techniques: Binding Assay, Sequencing

Journal: bioRxiv

Article Title: An improved TEAD dominant-negative protein inhibitor to study Hippo YAP1/TAZ-dependent transcription

doi: 10.1101/2024.10.03.615022

Figure Lengend Snippet: A . Western blot showing the expression of TEAD inhibitors in HEK293 cells 24hs after transfection. Unless otherwise indicated, TEADiv1 and TEADiv2 refer to the GFP-tagged version. B . Western blot of coimmunoprecipitation experiments showing that TEADiv2 reduces TEAD interaction with YAP1 and TAZ. C-D . Dose curve comparing TEADiv1 and TEADiv2 TEAD inhibition with 8x-TEAD-Luc reporter in HEK293 cells overexpressing YAP1 (C) or TAZ (D). n=3 independent experiments with technical duplicates. ns=p>0.05, **=p<0.005, paired t test. Mean ± SEM is shown. E . Dose curve comparing TEADimin TEAD inhibition with an 8x-TEAD-Luc reporter in HEK293 cells overexpressing YAP1 or TAZ. n=3 independent experiments with technical duplicates. Mean ± SEM is shown.

Article Snippet: Protein visualization was performed using the following antibodies: anti-GAPDH (Cell Signaling; clone no. 14C10; catalogue no. 2118; 1:2000), anti-GFP (Cell Signaling; clone no. D5.1; catalog no. 2956; 1:2000), anti-HA tag antibody (Cell Signaling; clone no. C29F4; catalog no. 3724; 1:1000), YAP1 (Cell Signaling; clone no. D8H1X; catalog no. 14074; 1:1000), TAZ (Cell Signaling; clone no. V386; catalog no. 4883; 1:1000), CYR61 (Cell Signaling; clone no. D4H5D; catalog no. 14479; 1:1000), and Pan-TEAD (Cell Signaling; clone no. D3F7L; catalog no. 13295; 1:1000).

Techniques: Western Blot, Expressing, Transfection, Inhibition

Journal: bioRxiv

Article Title: An improved TEAD dominant-negative protein inhibitor to study Hippo YAP1/TAZ-dependent transcription

doi: 10.1101/2024.10.03.615022

Figure Lengend Snippet: A-B . Graphs showing TEAD activity inhibition by TEADiv2. Transcriptional activity of TEAD was measured by luciferase assay in HEK293 cells transduced with TEADiv2 or GFP, each corresponding TEAD fused to the Gal4 DNA binding domain, UAS-luciferase, and either YAP1 (A) or TAZ (B). TEAD activity is reported as the fold decrease in TEADi compared with GFP control (dotted line indicates activity in GFP condition). n=3 independent experiments with technical duplicates. **=p<0.005, ***=p<0.001, ****=p<0.0001, t test comparing GFP vs TEADi. Mean ± SEM is shown. C . Comparison of GFP-tagged and HA-tagged TEADiv2 with 8x-TEAD-Luc reporter in HEK293 cells overexpressing YAP1 or TAZ. TEAD activity is reported as percentage inhibition over the activity of cells transfected with GFP instead of TEADi. n=3 independent experiments with technical duplicates. ns=p>0.05, t test. Mean ± SEM is shown.

Article Snippet: Protein visualization was performed using the following antibodies: anti-GAPDH (Cell Signaling; clone no. 14C10; catalogue no. 2118; 1:2000), anti-GFP (Cell Signaling; clone no. D5.1; catalog no. 2956; 1:2000), anti-HA tag antibody (Cell Signaling; clone no. C29F4; catalog no. 3724; 1:1000), YAP1 (Cell Signaling; clone no. D8H1X; catalog no. 14074; 1:1000), TAZ (Cell Signaling; clone no. V386; catalog no. 4883; 1:1000), CYR61 (Cell Signaling; clone no. D4H5D; catalog no. 14479; 1:1000), and Pan-TEAD (Cell Signaling; clone no. D3F7L; catalog no. 13295; 1:1000).

Techniques: Activity Assay, Inhibition, Luciferase, Transduction, Binding Assay, Control, Comparison, Transfection

Journal: Scientific Reports

Article Title: Influenza induces lung lymphangiogenesis independent of YAP/TAZ activity in lymphatic endothelial cells

doi: 10.1038/s41598-024-72115-6

Figure Lengend Snippet: List of antibodies used for histologic staining.

Article Snippet: Rabbit ⍺-TAZ (E8E9G) , Immunostaining , Cell Signaling Technologies , 83669 , 1:100.

Techniques: Staining, Immunostaining