protein 1  (Cell Signaling Technology Inc)

Journal: Genes & Diseases

Article Title: Restriction of YWHAB-mediated YAP cytoplasmic retention is a novel mechanism underlying stemness maintenance and chemoresistance in ovarian cancer peritoneal metastasis

doi: 10.1016/j.gendis.2025.101519

Figure Lengend Snippet: YWHAB depletion promotes YAP activity in OCPM. (A) The mRNA levels of YAP in YWHAB-knockdown CR primary OCPM and OVCAR3 cells and their control cells were measured by quantitative reverse-transcription PCR assay (one-way ANOVA; n = 3 biological replicates). (B) The protein levels of whole cell YAP and phosphorylated YAP in YWHAB-knockdown and control cells were measured by Western blot assay (one-way ANOVA; n = 3 biological replicates). (C, D) The protein levels of cytoplasmic (C) and nuclear (D) YAP in indicated cells were measured by Western blot (one-way ANOVA; n = 3 biological replicates). (E) The heatmap representing the relative mRNA levels of indicated YAP target genes in YWHAB-knockdown and control cells. (F) The transcriptional activities of YAP in indicated cells were examined by luciferase reporter assay (one-way ANOVA; n = 3 biological replicates). (G) The abundances of YAP and pan-TEAD immunoprecipitated by pan-TEAD antibody in indicated cells were examined by Western blot (one-way ANOVA; n = 3 biological replicates). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: The following antibodies were used to develop antibody-linked beads: Pan-TEAD (#13295, Cell Signaling Technology) and YAP (#14074, Cell Signaling Technology).

Techniques: Activity Assay, Knockdown, Control, Reverse Transcription, Western Blot, Luciferase, Reporter Assay, Immunoprecipitation

Journal: bioRxiv

Article Title: A PKM2-YAP reciprocal repartitioning modulates invasion of breast cancer cells

doi: 10.1101/2025.07.10.664113

Figure Lengend Snippet: (A) Representative confocal photomicrographs of breast cancer MDA-MB-231 cells grown on 0.4 kPa (top row of micrographs) and 20 kPa (bottom row of micrographs) polyacrylamide hydrogels coated with Collagen I and observed after 24□h of culture with staining for DNA (white using DAPI, left panel) and YAP (red, right panel). Yellow dotted areas represent nuclear area (n□= 3). Scale bar: 10□μm. Adjacent scatter plot graph depicting nucleus/cytoplasm ratio of YAP between MDA-MB-231 grown on 0.4 kPa and 20 kPa hydrogels (n = 3, n > 50). (B) Representative confocal photomicrographs of breast cancer MDA-MB-231 cells untreated (top row of micrographs) or treated with 40□μM TEPP-46, PKM2 cytoplasmic activator (second row from top) grown on 0.4 kPa (soft) hydrogels coated with Collagen I and observed after 24□h of culture with staining by□DNA (white using DAPI, left panel) and YAP (Red, right panel). Yellow dotted areas represent nuclear area (n□= 3). Scale bar: 10□μm. Adjacent scatter plot graph depicting nucleus/cytoplasm ratio of YAP between untreated and 40□μM TEPP-46 treated MDA-MB-231 cells on 0.4 kPa hydrogels (n = 3, N > 25). (C) Schematic depiction of YAP localization after PKM2 activation in soft gels. Error bars denote mean□±□SEM. Unpaired Student’s□ t □test was performed for statistical significance (* P □≤ 0.05, ** P □≤ 0.01, *** P □< 0.001, **** P □< 0.0001).

Article Snippet: The primary antibodies used in our studies are against PKM2 (4053S, CST), YAP (14074S, CST).

Techniques: Staining, Activation Assay

Journal: bioRxiv

Article Title: A PKM2-YAP reciprocal repartitioning modulates invasion of breast cancer cells

doi: 10.1101/2025.07.10.664113

Figure Lengend Snippet: (A) Representative confocal photomicrographs of breast cancer MDA-MB-231 cells untreated (top row of micrographs) or treated with 5□μM Verteporfin, YAP signalling inhibitor (second row from top) grown on 20 kPa (stiff) hydrogels coated with Collagen I and observed after 24□h of culture with staining by□DNA (white using DAPI, left panel) and F-actin (green using phalloidin, right panel (n□= 3). Scale bar: 20□μm. Graphs showing total cell area (top) and aspect ratio (second from top) of individual MDA-MB-231 cells untreated and treated with 5□μM Verteporfin on 20 kPa hydrogels (n□= 3, N□= 30 single cells). (B) Representative confocal photomicrographs of MDA-MB-231 cells untreated (top row of micrographs) or treated with 5□μM Verteporfin (second row from top) grown on 20 kPa (stiff) hydrogels coated with Collagen I and observed after 24□h of culture with staining by□DNA (white using DAPI, left panel) and PKM2 (Red, right panel) and adjacent scatter plot graph (right) depicting nucleus/cytoplasm ratio of PKM2 between untreated and 5□μM Verteporfin treated MDA-MB-231 cells on 20 kPa hydrogels (n=3, N=50). (n□= 3). Scale bar: 20□μm. (C) Epifluorescence photomicrographs of MDA-MB-231 cells with their migration tracks for 3 hours, control (left) and treated with 5□μM Verteporfin (right) on 20 kPa hydrogels and scatter plot graph (right) depicting migration speed (n=3, N > 20). See also Video S7, S8. (D) Graph measuring fold change of intracellular pyruvate (first from left), intracellular lactate (second from left), extracellular pyruvate (third from left), extracellular lactate (last from left) between untreated and 5□μM Verteporfin treated MDA-MB-231 cells on 20 kPa hydrogels (n=3). (E) Schematic depiction of PKM2-YAP reciprocal repartitioning. Error bars denote mean□±□SEM. Unpaired Student’s□ t □test was performed for statistical significance (* P □≤ 0.05, ** P □≤ 0.01, *** P □< 0.001, **** P □< 0.0001).

Article Snippet: The primary antibodies used in our studies are against PKM2 (4053S, CST), YAP (14074S, CST).

Techniques: Staining, Migration, Control