yap activator py 60  (MedChemExpress)

Journal: Biomolecules

Article Title: Mitophagy Activation via the YAP/Parkin Pathway Underlies the Neuroprotective Action of Tetramethylpyrazine in Cerebral Ischemia/Reperfusion Injury

doi: 10.3390/biom16030429

Figure Lengend Snippet: TMP modulates YAP activity and nuclear localization after cerebral ischemia/reperfusion. ( A ) Volcano plot showing differentially expressed genes following TMP treatment. Significant DEGs were defined as |log2FC| ≥ 1 and FDR < 0.05. ( B ) KEGG pathway enrichment analysis of differentially expressed genes. ( C ) Representative Western blot images of total YAP and p-YAP (Ser127) expression in Sham, MCAO/R, and TMP-treated (40 mg/kg) groups. ( D – F ) Quantitative analysis of p-YAP/β-actin, total YAP/β-actin, and p-YAP/YAP ratios. n = 3 per group. ( G ) Quantification of the YAP nuclear/cytoplasmic ratio. n = 4 per group. ( H ) Quantification of the percentage of cells exhibiting nuclear YAP localization. n = 4 per group. ( I ) Representative immunofluorescence images showing YAP (red), β-Tubulin (green), and DAPI (blue). Scale bars = 20 μm and 5 μm. ( J – L ) Correlation analyses between YAP nuclear localization and Parkin recruitment to mitochondria, LC3/TOM20 colocalization, and NeuN + neuronal density. Data are presented as mean ± SEM. ns, not significant; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Mitophagy and YAP activity were inhibited by Mdivi-1 (25 μM; MedChemExpress, Monmouth Junction, NJ, USA; catalog no. HY-15886) and Verteporfin (10 μM), respectively, applied 6 h before OGD and maintained during reoxygenation.

Techniques: Activity Assay, Western Blot, Expressing, Immunofluorescence

Journal: Biomolecules

Article Title: Mitophagy Activation via the YAP/Parkin Pathway Underlies the Neuroprotective Action of Tetramethylpyrazine in Cerebral Ischemia/Reperfusion Injury

doi: 10.3390/biom16030429

Figure Lengend Snippet: YAP activity is required for TMP-induced Parkin upregulation and mitophagy activation. ( A ) Representative immunofluorescence images showing LC3 (red), TOM20 (green), and nuclei (DAPI, blue) in the cortical penumbra. Scale bars: 20 μm and 5 μm (enlarged images). ( B ) Line-scan intensity profile analysis of LC3 and TOM20 signals along the indicated lines in enlarged images. ( C ) Quantification of LC3-positive mitochondria per mm 2 in different treatment groups. n = 5 per group. ( D , E ) Representative Western blot images and quantitative analysis of LC3-II and p62 protein levels normalized to β-actin. n = 3–4 per group. ( F ) Quantitative PCR analysis of Park2 mRNA expression in Sham, MCAO/R, TMP-treated (40 mg/kg), and TMP + VP groups. n = 3 per group. ( G , H ) Representative Western blot images and quantitative analysis of Parkin protein expression, with β-actin as a loading control. n = 3 per group. ( I ) Representative immunofluorescence images showing LC3 (red), TOM20 (green), and DAPI (blue) in OGD/R-injured neurons treated with TMP (100 μM), TMP + VP, TMP + VP + pCDH-Vector, or TMP + VP + pCDH- Park2 . Scale bars: 20 μm. ( J ) Quantification of LC3 recruitment to mitochondria based on mean fluorescence intensity (MFI). n = 3 independent experiments. Data are presented as mean ± SEM. ns, not significant; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Original images of Western blotting can be found in .

Article Snippet: Mitophagy and YAP activity were inhibited by Mdivi-1 (25 μM; MedChemExpress, Monmouth Junction, NJ, USA; catalog no. HY-15886) and Verteporfin (10 μM), respectively, applied 6 h before OGD and maintained during reoxygenation.

Techniques: Activity Assay, Activation Assay, Immunofluorescence, Western Blot, Real-time Polymerase Chain Reaction, Expressing, Control, Plasmid Preparation, Fluorescence

Journal: bioRxiv

Article Title: Ionic Regulation of Mechanosurveillance and Metastasis via the MRTFA/KCNMB1 Axis

doi: 10.64898/2026.01.13.699089

Figure Lengend Snippet: (A) Venn diagram and gene ontology analyses of the SRF independent genes that are exclusively upregulated by MRTFA Y238A in both AT-3 and E0771 cells to at least 2-fold or more. See Table S2 for full results. (B) Examples of statistically significant GSEA for MRTFA high/low breast cancer patient tumors from TCGA. (C) Examples of statistically significant GSEA for SRF high/low breast cancer patient tumors from TCGA. (D) Relative mRNA expression levels of MRTFA and MRTFB in shMRTFA/B and SRF in shSRF mouse cell lines. Data are shown as fold change calculated from 3 technical replicates per sample and normalized to GAPDH levels. Each graph is representative of 3 independent experiments. (E) Relative mRNA expression levels of MRTFA and MRTFB in shMRTFA/B and SRF in shSRF human cell lines. Data are shown as fold change calculated from 3 technical replicates per sample and normalized to GAPDH levels. Each graph is representative of 3 independent experiments. (F) Relative mRNA expression levels of canonical YAP target gene CCN2, which encodes connective tissue growth factor (CTGF), and KCNMB1 in AT-3 cells overexpressing wildtype YAP or hyperactive YAP 5SA mutant or treated for 24 hours with TGFβ inhibitor SB-505124. (G) KCNMB1 human and mouse genetic loci analyzed for SRF (purple tracks) binding by using University of California Santa Cruz Genome Browser data publicly available Chromatin immunoprecipitation sequencing data. Each purple solid line under the ReMap density map indicates an independent dataset where SRF was found to be associated with the locus. In the Encode cCREs track, red solid boxes indicate promoter regions, orange and yellow solid boxes indicate proximal and distal enhancers, respectively, and blue tracks indicate CTCF only binding regions. (H) Chromatin immunoprecipitation-PCR from AT-3 cells using KCNMB1 primers and the indicated antibodies against SRF, MRTFA, histone H3 (as a positive control) and IgG (negative control). Water was used as input material as a negative control for the PCR reaction. Image is representative of 3 independent experiments. (I) Raw counts from bulk RNA-seq data for Myocd , Mrtfa , and Mrtfb genes in AT-3 and E0771 cells.

Article Snippet: To over express YAP (Addgene #33091) and its active mutant YAP 5SA (Addgene #33093) were cloned into pLVX-puro (Takara Bio Catalog No. #632164).

Techniques: Expressing, Mutagenesis, Binding Assay, ChIP-sequencing, Chromatin Immunoprecipitation, Positive Control, Negative Control, RNA Sequencing