pak2 inhibitor frax597  (TargetMol)

Journal: Breast Cancer Research : BCR

Article Title: PAK2 promotes CTC cluster formation by phosphorylating E-cadherin to enhance cell-cell adhesion in breast cancer

doi: 10.1186/s13058-025-02199-z

Figure Lengend Snippet: Protein-protein interaction network analysis, hub gene identification, and prognostic significance of PAK2 in breast cancer. a PPI network constructed for 193 coDEGs. b Top 30 hub genes identified using the CytoHubba plugin in Cytoscape based on MCC scores. c Venn diagram highlighting the selection of PAK2 as a hub gene. d , e Representative IHC staining images of PAK2 expression in normal breast tissue and breast cancer tissues from the Human Protein Atlas database. f , g Kaplan-Meier analysis of the prognostic impact of PAK2 expression on DMFS in breast cancer patients

Article Snippet: The PAK2 inhibitor FRAX597 was purchased from TargetMol (Shanghai, China).

Techniques: Construct, Selection, Immunohistochemistry, Expressing

Journal: Breast Cancer Research : BCR

Article Title: PAK2 promotes CTC cluster formation by phosphorylating E-cadherin to enhance cell-cell adhesion in breast cancer

doi: 10.1186/s13058-025-02199-z

Figure Lengend Snippet: PAK2 expression and cluster formation in breast cancer cell lines. a mRNA expression levels of PAK2 in breast cancer cell lines and the normal mammary epithelial cell line MCF10A. b – f Quantification of cell cluster formation in four breast cancer cell lines under suspension culture conditions at 12, 24, and 48 h. g Immunoblot analysis of total PAK2 and phospho-PAK2 (Ser141) levels in BT549, MDA-MB-231, SKBR3, and AU565 breast cancer cells. SKBR3 cells were used as the reference (set to 1) for relative expression

Article Snippet: The PAK2 inhibitor FRAX597 was purchased from TargetMol (Shanghai, China).

Techniques: Expressing, Suspension, Western Blot

Journal: Breast Cancer Research : BCR

Article Title: PAK2 promotes CTC cluster formation by phosphorylating E-cadherin to enhance cell-cell adhesion in breast cancer

doi: 10.1186/s13058-025-02199-z

Figure Lengend Snippet: PAK2 promotes cell cluster formation. a , b Immunoblot analysis showing PAK2 expression in BT549 cells after overexpression or knockdown. c Representative images and quantification of cluster formation in BT549 cells transfected with control vector (Con) or lenti-PAK2 (OE-PAK2). d Representative images and quantification of cluster formation in BT549 cells transfected with control siRNA (siCon) or siPAK2-1

Article Snippet: The PAK2 inhibitor FRAX597 was purchased from TargetMol (Shanghai, China).

Techniques: Western Blot, Expressing, Over Expression, Knockdown, Transfection, Control, Plasmid Preparation

Journal: Breast Cancer Research : BCR

Article Title: PAK2 promotes CTC cluster formation by phosphorylating E-cadherin to enhance cell-cell adhesion in breast cancer

doi: 10.1186/s13058-025-02199-z

Figure Lengend Snippet: Effects of PAK2 on breast cancer cell progression. a , b Colony formation assays performed in BT549 cells with PAK2 overexpression or knockdown. c , d Wound healing assays evaluating the migration of BT549 cells with PAK2 overexpression or knockdown. e Transwell assays evaluating the invasive capacity of BT549 cells with PAK2 overexpression or knockdown

Article Snippet: The PAK2 inhibitor FRAX597 was purchased from TargetMol (Shanghai, China).

Techniques: Over Expression, Knockdown, Migration

Journal: Breast Cancer Research : BCR

Article Title: PAK2 promotes CTC cluster formation by phosphorylating E-cadherin to enhance cell-cell adhesion in breast cancer

doi: 10.1186/s13058-025-02199-z

Figure Lengend Snippet: PAK2 mediates cell cluster formation via E-cadherin phosphorylation. a , b GSEA showing the association between PAK2 and the adherens junction pathway. c , d Immunoblot analysis of E-cadherin and phosphorylated E-cadherin (Ser840) in BT549 cells after PAK2 overexpression or knockdown

Article Snippet: The PAK2 inhibitor FRAX597 was purchased from TargetMol (Shanghai, China).

Techniques: Phospho-proteomics, Western Blot, Over Expression, Knockdown

Journal: Breast Cancer Research : BCR

Article Title: PAK2 promotes CTC cluster formation by phosphorylating E-cadherin to enhance cell-cell adhesion in breast cancer

doi: 10.1186/s13058-025-02199-z

Figure Lengend Snippet: PAK2 interacts and colocalizes with E-cadherin. a Protein docking map and binding site of PAK2 for E-cadherin. The yellow color represents PAK2, and the blue color represents E-cadherin. b IF staining illustrating the colocalization of PAK2 and E-cadherin in MCF10A and BT549. c Co-IP analysis showing the association between PAK2 and E-cadherin in breast cancer cells

Article Snippet: The PAK2 inhibitor FRAX597 was purchased from TargetMol (Shanghai, China).

Techniques: Binding Assay, Staining, Co-Immunoprecipitation Assay

Journal: Breast Cancer Research : BCR

Article Title: PAK2 promotes CTC cluster formation by phosphorylating E-cadherin to enhance cell-cell adhesion in breast cancer

doi: 10.1186/s13058-025-02199-z

Figure Lengend Snippet: FRAX597 inhibits PAK2 activity and CTC cluster formation. a CCK-8 assay evaluating the viability of BT549 after 48 h treatment with FRAX597. b Immunoblot analysis of total PAK2 and phosphorylated PAK2 protein levels in BT549 treated with 2.5 or 5 µM FRAX597 for 24 and 48 h. c Representative images and quantification of BT549 cell aggregation after 24 h FRAX597 treatment. d Immunoblot analysis of total and phosphorylated PAK2, and total and phosphorylated E-cadherin in BT549 treated with 2.5 or 5 µM FRAX597 for 24 h

Article Snippet: The PAK2 inhibitor FRAX597 was purchased from TargetMol (Shanghai, China).

Techniques: Activity Assay, CCK-8 Assay, Western Blot

Journal: Breast Cancer Research : BCR

Article Title: PAK2 promotes CTC cluster formation by phosphorylating E-cadherin to enhance cell-cell adhesion in breast cancer

doi: 10.1186/s13058-025-02199-z

Figure Lengend Snippet: FRAX597 suppresses breast cancer progression. a – c Effects of FRAX597 on BT549 cell proliferation, migration and invasion. d Schematic diagram of in vivo experimental design. e , f Tumor volume, tumor weight and body weight measurements of nude mice treated with FRAX597

Article Snippet: The PAK2 inhibitor FRAX597 was purchased from TargetMol (Shanghai, China).

Techniques: Migration, In Vivo

Journal: Breast Cancer Research : BCR

Article Title: PAK2 promotes CTC cluster formation by phosphorylating E-cadherin to enhance cell-cell adhesion in breast cancer

doi: 10.1186/s13058-025-02199-z

Figure Lengend Snippet: FRAX597 reduces breast cancer metastasis in vivo. a Representative H&E staining images of lung tissue sections. a – c Quantification of the number and size of metastatic nodules in the lungs. d Representative images of three phenotypes of CTCs detected in mouse blood. e – g Quantification of the number and percentage of single CTCs and CTC clusters in both groups

Article Snippet: The PAK2 inhibitor FRAX597 was purchased from TargetMol (Shanghai, China).

Techniques: In Vivo, Staining

Journal: Breast Cancer Research : BCR

Article Title: PAK2 promotes CTC cluster formation by phosphorylating E-cadherin to enhance cell-cell adhesion in breast cancer

doi: 10.1186/s13058-025-02199-z

Figure Lengend Snippet: FRAX597 decreases the proportion of E/M-CTCs ( a ) Ratios of E-CTC, M-CTC and E/M-CTC in the control group. b Ratios of E-CTC, M-CTC and E/M-CTC in the FRAX597 treatment group

Article Snippet: The PAK2 inhibitor FRAX597 was purchased from TargetMol (Shanghai, China).

Techniques: Control

Journal: Breast Cancer Research : BCR

Article Title: PAK2 promotes CTC cluster formation by phosphorylating E-cadherin to enhance cell-cell adhesion in breast cancer

doi: 10.1186/s13058-025-02199-z

Figure Lengend Snippet: Schematic diagram. PAK2-mediated phosphorylation of E-cadherin at Ser840 promotes CTC clustering and metastasis

Article Snippet: The PAK2 inhibitor FRAX597 was purchased from TargetMol (Shanghai, China).

Techniques: Phospho-proteomics

Journal: International journal of biological sciences

Article Title: Neutrophil Extracellular Traps Delay Diabetic Wound Healing by Inducing Endothelial-to-Mesenchymal Transition via the Hippo pathway.

doi: 10.7150/ijbs.78046

Figure Lengend Snippet: Figure 3. NETs phosphorylate Merlin/NF2 and suppress the Hippo-YAP pathway in EC. (A) Western blotting analysis of YAP, p-YAP, Last1, Mst1, and MOB1. β-actin is used as a loading control. (B) Immunofluorescence images showing localization and expression of TLR-9, PAK2, and Merlin/NF2 by triple dyeing. Scare bar = 20μm. (C) Western blotting analysis of PAK2, Merlin/NF2, and p-Merlin (Ser518). β-actin is used as a loading control. (D) Western blotting analysis of Merlin/NF2, p-Merlin (Ser518), YAP, and p-YAP. β-actin is used as a loading control. (E) Immunoblotting of HUVECs that were infected with lentivirus expressing the indicated flagMerWT constructs and immunoprecipitated with anti-flag antibody to detect the binding of endogenous PAK2 to flagMerWT constructs. In the control group, cells were infected with lentiviruses expressing empty vector. (F) Western blotting analysis of Merlin/NF2, YAP, and p-YAP. β-actin is used as a loading control. All the cells were infected with lentiviruses expressing shRNA of NF2. At 3 days post-lentivirus infection, cells were infected with lentiviruses expressing empty mCherry vector (Empty), flagMerWT, flagMerSA, or flagMerSD. Analysis was performed at 3 days post-lentivirus infection. FRAX597 is an ATP-competitive inhibitor of PAK2. The flagMerWT expresses the wild-type form of Merlin/NF2. The flagMerSA constitutively expresses a growth-inhibitory (activated) form of Merlin/NF2 in which there is an alanine substitution at the S518 phosphorylation site. The flagMerSD expresses a phosphomimetic (inhibited) form of Merlin/NF2. *P < 0.05, **P < 0.01, the variables between two groups were compared using Student’s t test. For variables of more than two groups, statistical analysis was performed by one-way ANOVA followed by the SNK-q post hoc test. Data are shown as the mean ± SD, n = 6 in each group in this figure.

Article Snippet: The cells were then incubated with neutrophils (control group), NETs (NETs group), NETs treated with 0.1 mg/ml DNase I (InvivoGen, San Diego, CA, USA), NETs and TLR9 antagonist (ODN 2088, San Diego, CA, USA), NETs and PAK2 inhibitor (FRAX597, Selleck Chemicals, Houston, TX, USA), and NETs and Hippo pathway inhibitor (XMU-MP-1, Selleck Chemicals, Houston, TX, USA) for 12 h at 37°C under 5% CO2.

Techniques: Western Blot, Control, Immunofluorescence, Expressing, Infection, Construct, Immunoprecipitation, Binding Assay, Plasmid Preparation, shRNA, Phospho-proteomics

Journal: International journal of biological sciences

Article Title: Neutrophil Extracellular Traps Delay Diabetic Wound Healing by Inducing Endothelial-to-Mesenchymal Transition via the Hippo pathway.

doi: 10.7150/ijbs.78046

Figure Lengend Snippet: Figure 7. Hypothetical model of NET participation in delayed healing of diabetic wounds by inducing endothelial-to-mesenchymal transition via the Hippo-YAP pathway. Diabetic wound environment primes neutrophils to form NETs. NETs induce the activation of PAK2 via the membrane receptor TLR-9 in ECs. PAK2 phosphorylates the intracellular protein Merlin/NF2 to inhibit the Hippo-YAP signaling pathway. YAP binds to the transcription factor SMAD2. Then they translocate from the cytoplasm into the nucleus together to further induce the endothelial-to-mesenchymal transformation, which impedes angiogenesis and delays wound healing.

Article Snippet: The cells were then incubated with neutrophils (control group), NETs (NETs group), NETs treated with 0.1 mg/ml DNase I (InvivoGen, San Diego, CA, USA), NETs and TLR9 antagonist (ODN 2088, San Diego, CA, USA), NETs and PAK2 inhibitor (FRAX597, Selleck Chemicals, Houston, TX, USA), and NETs and Hippo pathway inhibitor (XMU-MP-1, Selleck Chemicals, Houston, TX, USA) for 12 h at 37°C under 5% CO2.

Techniques: Activation Assay, Membrane, Transformation Assay

Journal: The American Journal of Pathology

Article Title: Neutrophil-Derived Reactive Oxygen Orchestrates Epithelial Cell Signaling Events during Intestinal Repair

doi: 10.1016/j.ajpath.2019.07.017

Figure Lengend Snippet: Isotope-coded affinity tags identify p21-activated kinase 2 (Pak2) as a mediator of oxidative signaling. A: Biotin-iodoacetamide (BIAM) labeling analysis of Pak2 from SK-CO-15 cells treated with 1 mmol/L H2O2 for 15 minutes. Immunoblot analysis was used to detect the levels of total Pak2. Immunoblot shown is representative of at least three replicate experiments. B: Immunofluorescence analysis of SK-CO-15 cells treated with the Pak2 inhibitor FRAX597 at 2 μmol/L for 1 hour, followed by exposure to 1 mmol/L H2O2 for 15 minutes. Analysis detected phosphorylated Crk-associated substrate (p-Cas; red), and total focal adhesion kinase (FAK; green) localization in nuclei was stained with DAPI (blue). Scale bars = 20 μm (B). CTL, control.

Article Snippet: FAK inhibitor PF-562271 was used at 500 nmol/L final concentration, and the p21-activated kinase 2 (PAK2) inhibitor FRAX597 (Selleckchem, Houston, TX) was used at 2.5 μmol/L final concentration.

Techniques: Labeling, Western Blot, Immunofluorescence, Staining