inhibition  (MedChemExpress)

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: bioRxiv

Article Title: A defined 2D system for generating and expanding human basal radial glia from iPSCs

doi: 10.64898/2026.04.10.714524

Figure Lengend Snippet: A . Representative immunostainings of Stage-2 cultures treated with candidate niche factors in addition to FGF2. Cells were stained for phosphorylated vimentin (pVIM) to label mitotic progenitors and GFAP to visualize RG-like morphology. Images are shown for two independent iPSC lines. Scale bars, 50 µm. B . Quantification of proliferative cells (pVIM + ) in Stage-2 cultures treated with candidate niche factors during the initial niche-factor screening. Data represent two batches from two independent iPSC lines (327,413 total cells quantified). C . Representative brightfield images illustrating the effects of selected pathway modulators on Stage-2 RG cultures. Stage-2 RG cultures maintained with FGF2 alone, or supplemented for 3 weeks with EGF or the YAP activator TRULI. Both conditions resulted in altered culture morphology and reduced long-term stability. Scale bars, 50 µm. D–E . Quantification of proliferative cells (D; pVIM+ fraction) and average Nestin+ process length (E) in Stage-2 cultures treated with individual candidate factors or the final factor combination. Each point represents the mean value of one biological replicate. Circles and triangles indicate the two independent iPSC lines used. Data were obtained across five (pVIM) or six (Nestin) independent differentiation batches. Boxplots display the median (centre line) and interquartile range (box; 25th–75th percentiles), with whiskers extending to 1.5× the interquartile range. Statistical comparisons were performed on replicate means using Wilcoxon rank-sum tests against the FGF condition, with Benjamini–Hochberg correction applied for multiple comparisons. F . Quantification and representative time-lapse imaging of IST in Stage-3 bRG cultures. Each dot represents the mean fraction of cells undergoing IST per independent differentiation batch (2 lines, 3 batches each). Data are shown as mean ± SD. Representative time-lapse images illustrate IST progression at the indicated time points. Yellow asterisks mark cells undergoing IST. G . KEGG pathway enrichment analysis comparing Stage-3 and Stage-2 cultures, highlighting pathways associated with extracellular matrix interactions, integrin signaling, cytoskeletal regulation and neuroactive ligand-receptor signaling. H . Quantification of average speed in Stage-3 bRG cultures following treatment with different concentrations of the PAK2 inhibitor FRAX597. Speed was assessed during 96 h live-cell imaging with 10 min acquisition intervals. For each independent replicate (n=3), the mean speed was calculated across all tracked cells. Each point represents one independent replicate, defined as a unique combination of cell line and experimental batch. Data were obtained from two independent cell lines across three independent differentiation batches. Data is reported as mean ± SD. Statistical comparisons against the control condition were performed on replicate means using paired Wilcoxon signed-rank tests, with Benjamini-Hochberg correction applied for multiple comparisons, only p.adj < 0.05 are reported.

Article Snippet: For pharmacological inhibition, FRAX597 (MedChemExpress) was added 24 h prior to imaging at the indicated concentrations.

Techniques: Staining, Imaging, Live Cell Imaging, Control

Journal: bioRxiv

Article Title: Phosphorylation Protects Oncogenic RAS from LZTR1-Mediated Degradation

doi: 10.64898/2026.01.07.698128

Figure Lengend Snippet: A ) Schematic for a putative kinase phosphorylating RAS T148. B ) Scatter plot of kinase-related genes from the KRAS stability screens in . C ) Venn diagram of kinase genes with decreased KRAS expression (≥0.5 KRAS stability score), RAS interaction partners from BioID2 studies (≥0.8 log2 fold enrichment, ), and essential genes identified in RAS-dependent MM lines (CSS<-1.0, 19 ). D ) Western blot analysis of RAS expression following treatment with 100 nM of the indicated drugs for 24 and 48 hours in XG7, RPMI 8226, and SKMM1 MM cells, n=5. E ) Evaluation of pT148 RAS phosphorylation in cells treated with MG-132 for 8 hours before lysis and the indicated drugs for 48 hours, n=3. F ) Co-IP of PAK1 and PAK2 with ectopically expressed mNG-KRAS G12D in the indicated MM lines, n=2. G ). Indicated cells expressing listed shRNAs or DN PP1C were treated with a dose titration of RMC-6236. Data shown is the ratio of experiment (listed shRNA or DN PP1C) versus shCTRL. Values shown are the average of 2 experiments. H ) in vitro kinase assay of recombinant PAK1 on recombinant KRAS as a substrate. Western blot analysis of phospho-T148 RAS was used as a readout. Inhibition with FRAX597 was used to demonstrate specificity, n=4. I ) Cell viability in XG7 cells ectopically expressing KRAS G12D or KRAS G12D+T148D and treated with indicated drugs: either 100 nM FRAX527 and/or 8 nM RMC-6236. J ) Tumor volume for MM.1S xenografts treated with vehicle (gray), 3 mg/kg FRAX597 (purple), 5 mg/kg RMC-6236 (pink), or the combination of both drugs (red). K ) Inhibition with FRAX597 primes RAS for degradation.

Article Snippet: FRAX597, RMC-6236 (SelleckChem) were dissolved in DMSO and diluted in equal volumes at the specified concentrations.

Techniques: Expressing, Western Blot, Phospho-proteomics, Lysis, Co-Immunoprecipitation Assay, Titration, shRNA, In Vitro, Kinase Assay, Recombinant, Inhibition

Journal: bioRxiv

Article Title: PERK inhibition rewires translational and CMGC protein kinase networks into an antiviral state

doi: 10.1101/2025.10.08.681090

Figure Lengend Snippet: (A) P-peptides from the same samples described in were extracted by IMAC and analyzed by LC-MS/MS resulting in 12585 peptide IDs. The heatmap demonstrates mean changes of P-peptides, their sequences and the identified P-sites (in red) corresponding to the N or S proteins (green or yellow colors) and to host proteins c-Jun and p38 MAPK (MAPK14) for each of the six conditions analyzed. Data are from four biologically independent experiments and two technical replicates. (B) Volcano plots show mean Log 2 -fold changes of all P-peptides and their significance from four biologically independent experiments and two technical replicates. Numbers in brackets and red or blue colors indicate up- or down-regulated P-peptides in response to CoV or PERKi treatment (-Log 10 p value ≥ 1.3, student’s t-test). P-peptides corresponding to N or S protein and individual P-sites are highlighed in green or yellow, respectively. Red text indicates sets of P-peptides suppressed by PERKi treatment in infected cells. (C) Venn diagrams indicating the overlap of P-peptides differentially phosphorylated in response to CoV or PERKi based on (-Log 10 p value ≥ 1.3, student’s t-test). Numbers in brackets indicate the overlap of gene IDs corresponding to the identified P-peptides. (D) Overrepresentation analysis was performed side by side based on gene IDs corresponding to P- peptides for factors that were up- or down-regulated in response to PERKi after 12 h or 24 hpi. The Venn diagram shows the overlap of the top100 enriched pathway terms across all conditions. (E) Heatmap showing the top5 most strongly enriched pathway terms representing exclusively suppressed P-protein sets (upper table) or those with a mixed pattern of regulation in response to PERKi (lower table). Yellow color highlights the Rho GTPase pathway. (F) Scatter plots showing mean abundancies of the P-peptides that are suppressed by PERKi 24 hpi, segregated according to the pathways identified in (E), along with their values across all conditions. Data are from four biological replicates and two technical replicates, black lines indicate medians. Red colors mark P-peptides belonging to protein kinases. Blue colors highlight P-peptides of PAK2. (G-J) Huh7 cells were infected for 24 h with HCoV-229E (H, MOI=l) and treated with group I PAK inhibitors FRAX469 (G, I) or FRAX597 (H-J) at the indicated concentrations, thapsigargin (l µM), solvent (DMSO) or were left untreated. (G-H) Whole cell extracts were used to analyze phosphorylation and expression of proteins by Western blotting. One representative out of four biologically independent experiments is shown. (I-J) Quantification of the indicated (P)-proteins from four replicates. Shown are mean values ± s.d. including non-linear regression lines used to estimate IC 50 values. (K) Experiments were performed as described in (G-J), including two additional concentrations of PAKi (10 µM, 25 µM). Supernatants of cells were used to analyze viral titers by plaque assays and parallel cell cultures were used to assess cell viability. Left graphs show mean titers ± s.d. from four biologically independent experiments. The same data were used to estimate EC 50 values by non-linear regression (middle graphs). Right graphs show mean normalized cell viability values ± s.d. including non-linear regression lines to estimate CC 50 values as indicated, obtained from three biologically independent experiments. Asterisks indicate p values (*p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001) derived from Kruskal Wallis tests. (C-F) Amber rectangles denote the sets of P-peptides and their corresponding gene IDs suppressed significantly by PERKi at 24 hpi (-Log 10 p value ≥ 1.3, student’s t-test).

Article Snippet: FRAX486 (TGM6840) and FRAX597 (TGM6014) were obtained from TargetMol and KIRA6 (532281) was purchased from Calbiochem through Merck-Millipore.

Techniques: Liquid Chromatography with Mass Spectroscopy, Infection, Solvent, Phospho-proteomics, Expressing, Western Blot, Derivative Assay