β catenin inhibitor msab  (MedChemExpress)

Journal: bioRxiv

Article Title: A Wnt-responsive fibrocartilage progenitor system coordinates postnatal mandibular condylar cartilage growth

doi: 10.64898/2026.03.25.714159

Figure Lengend Snippet: Isolated Wnt-responsive cells were cultured in vitro and treated with the Wnt pathway inhibitor XAV-939 (1 μM or 5 μM), the Foxm1 inhibitor RCM-1 (4 μM or 20 μM), or the Wnt agonist Wnt agonist 1 (0.2 nM or 1 nM). Cell proliferation was quantified by measuring total cell number after treatment. Pharmacological inhibition of canonical Wnt signaling using XAV-939 significantly reduced cell expansion compared with control cultures. Similarly, inhibition of Foxm1 using RCM-1 markedly suppressed proliferation. In contrast, activation of canonical Wnt signaling by Wnt agonist 1 increased cell numbers relative to controls. These findings support a cooperative role for canonical Wnt signaling and Foxm1 in promoting proliferative capacity of Wnt-responsive cells in vitro . Bars represent mean ± s.d. Each dot represents one biologically independent sample. Statistical significance was determined by one-way ANOVA followed by multiple comparisons testing. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.

Article Snippet: Pharmacological treatments were performed using the Foxm1 inhibitor RCM-1, the β-catenin inhibitor XAV-939, or a canonical Wnt pathway activator (Wnt agonist 1) (all from Selleck Chemicals, Japan).

Techniques: Isolation, Cell Culture, In Vitro, Inhibition, Control, Activation Assay

Journal: Cells

Article Title: TMAO-Triggered Endothelial–Mesenchymal Transition and Microvesicle Release as Mediators of Vascular Smooth Muscle Cell Osteogenic Differentiation and Vascular Calcification

doi: 10.3390/cells15050466

Figure Lengend Snippet: β-catenin inhibition attenuates endothelial exosome-induced β-catenin activation in HAVSMCs. ( A , C ) Representative Western blot images showing non-phosphorylated (active) β-catenin protein expression in human aortic vascular smooth muscle cells (HAVSMCs) treated with endothelial cell-derived exosomes (EC-EXOs) obtained from TNFα-, TGFβ-, or TMAO-stimulated endothelial cells, in the presence or absence of the β-catenin transcriptional inhibitor ICG-001 for 8 days. β-actin was used as a loading control. ( B , D ) Quantitative densitometric analysis demonstrates a significant increase in β-catenin protein levels following EC-EXO treatment, which was markedly reduced upon β-catenin inhibition with ICG-001. Protein expression levels were normalized to β-actin and expressed as fold change relative to vehicle-treated controls. Data are presented as mean ± standard deviation (SD) from three independent biological replicates. Statistical significance was determined by one-way ANOVA followed by Tukey’s post hoc test to assess differences between EC-EXO treatment groups and the effect of β-catenin inhibition. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: To determine the requirement of Wnt/β-catenin signaling in endothelial exosome-induced osteogenic reprogramming, HAVSMCs were treated with the selective β-catenin/CBP transcriptional inhibitor ICG-001 (TargetMol).

Techniques: Inhibition, Activation Assay, Western Blot, Expressing, Derivative Assay, Control, Standard Deviation

Journal: Cells

Article Title: TMAO-Triggered Endothelial–Mesenchymal Transition and Microvesicle Release as Mediators of Vascular Smooth Muscle Cell Osteogenic Differentiation and Vascular Calcification

doi: 10.3390/cells15050466

Figure Lengend Snippet: β-catenin inhibition suppresses endothelial exosome-induced osteogenic gene expression in HAVSMCs. ( A – E ) Quantitative real-time PCR analysis of osteogenic gene expression in HAVSMCs treated with endothelial cell-derived exosomes (EC-EXOs) from TNFα-, TGFβ-, or TMAO-stimulated endothelial cells, in the presence of the β-catenin inhibitor ICG-001. Relative mRNA expression levels of ( A ) SM22A, ( B ) αSMA, ( C ) RUNX2, ( D ) osteopontin (OPN), and ( E ) tissue-nonspecific alkaline phosphatase (TNAP) were normalized to housekeeping genes and expressed relative to vehicle-treated control cells (0.1% v / v DMSO). EC-EXO co-treatment with ICG-001 significantly attenuated the expression of RUNX2, OPN, and TNAP, indicating that β-catenin signaling is required for endothelial exosome-induced osteogenic reprogramming of HAVSMCs. Data are presented as mean ± SD from three independent biological replicates. Statistical significance was assessed using one-way ANOVA, followed by post-hoc analysis. * p < 0.05, ** p < 0.01, *** p < 0.001, vs. CTL vehicle.

Article Snippet: To determine the requirement of Wnt/β-catenin signaling in endothelial exosome-induced osteogenic reprogramming, HAVSMCs were treated with the selective β-catenin/CBP transcriptional inhibitor ICG-001 (TargetMol).

Techniques: Inhibition, Gene Expression, Real-time Polymerase Chain Reaction, Derivative Assay, Expressing, Control

Journal: EMBO Reports

Article Title: Liver sinusoidal endothelial cells constitute a major route for hemoglobin clearance

doi: 10.1038/s44319-025-00673-5

Figure Lengend Snippet: ( A ) Cell-surface expression of the CD163 receptor on LSECs and KCs was measured by flow cytometry and presented as fluorescence intensity. ( B ) Uptake of free Hb or Hb in the presence of haptoglobin (Hp, Hb:Hp) in primary murine LSEC cultures was measured with flow cytometry. ( C ) Hb-AF647 vesicle localization was imaged with microscopy in NPCs in vitro cultures depleted of macrophages. Arrows indicate Hb-AF647 (blue) presence in the EEA1 + (green) and Actin + (white) vesicles. Areas in the highlighted rectangles are shown at a higher 5× magnification (left corner). Scale bars, 20 μm. Single-channel images for this figure are presented in Fig. . ( D ) Co-localization of Hb-AF647 (red) with rhodamine-dextran (green) was imaged with microscopy in STAB2 + LSECs (white); nuclei are stained in blue. The arrow indicates colocalization of Hb and rhodamine-dextran. The area in the highlighted rectangle is shown at a higher 5× magnification in separate channels below. Scale bar, 20 μm. Single-channel images for this figure are presented in Fig. . ( E ) EEA1 + vesicles (green) were imaged with microscopy in human NPCs cultures stained with the LSEC marker CD36 (orange); nuclei are stained in blue. ( F ) Texas red-dextran uptake (orange) was imaged with microscopy in human NPCs cultures stained with the LSEC marker CD36 (blue). ( G ) NPCs were pre-treated with the clathrin-mediated endocytosis inhibitor chlorpromazine (CPZ), the macropinocytosis blocker EIPA, actin polymerization blocker latrunculin A (LAT-A), inhibitor of Cdc42 GTPase ML141, β-catenin inhibitor PRI-724, and with micropinocytosis blocker nystatin before Hb-AF750 treatment for 10 min. Flow cytometry was used to determine Hb uptake by the LSECs. ( H ) Flow cytometric analysis of CD36 hi CD32B hi and CD36 mid CD32B mid human primary LSECs, treated with Hb-AF647 and exposed to EIPA, LAT-1, and CPZ. Numbers indicate MFI. Data are representative of three independent donors of human NPCs. ( I ) Mice were injected for 1 h with Hb-AF750 or albumin-AF750, both at 10 µg/mouse. The fluorescence intensity of AF750, the percentage of AF750 + LSECs, and KCs populations were measured with flow cytometry. ( J ) NPCs were pre-treated with CPZ, EIPA, LAT-A, ML141, and PRI-724 before albumin-AF647 treatment for 10 min. Flow cytometry was used to determine albumin uptake by the LSECs. ( C – F ) Scale bars, 20 μm. Numerical data are expressed as mean ± SEM, each data point represents one biological replicate (individual mouse or independent cell-based experiment), n = 3 ( A , B ), 3–6 ( G ), 4–5 ( I ), 4 ( J ). Welch’s unpaired t test was used to determine statistical significance in ( A , G ) for nystatin, two-way ANOVA with Tukey’s Multiple Comparison tests was used in ( I ); while one-way ANOVA with Tukey’s Multiple Comparison tests was used in ( G , J ). Exact P values are shown on graphs; values above bars in ( G , J ) indicate comparison with untreated controls. .

Article Snippet: The day after cell seeding, the cells were stimulated with freshly prepared fluorescently stained hemoglobin (Hb-AF647 or Hb-AF750, 0.5 μg/ml), hemoglobin-haptoglobin complex (Hb-AF750:Hp, 0.5 μg:15 μg/ml), Dextran Tetramethylrhodamine, 70,000 MW, Lysine Fixable (1 mg/ml, ThermoScientific, #D1818), Dextran, Texas Red, 70,000 MW, Neutral (0.25 mg/ml, ThermoScientific, # D1830) or Albumin from Bovine Serum (BSA-AF647) (1 μg/ml, ThermoScientific, # A34785 ) for 1 h. When indicated, the following compounds were used for pre-treatments: Wnt/beta-catenin inhibitor PRI-724 (5 μM, for 24 h, Selleckchem, #S8968); inhibitor of clathrin-mediated endocytosis, chlorpromazine (2 μM, for 1 h, Sigma-Aldrich, #C8138); inhibitor of macropinocytosis, EIPA (25 μM, for 1 h, MedChemExpress, #HY-101840); inhibitor of Cdc42 GTPase, ML141 (10 μM, for 1 h, Selleckchem, #S7686), an actin-depolymerizing agent, latrunculin A (1 μM, for 30 min, Tocris, #3973) and inhibitor of caveolae-mediated endocytosis, nystatin (240 U/ml for 30 min, Sigma-Aldrich, # N1638).

Techniques: Expressing, Flow Cytometry, Fluorescence, Microscopy, In Vitro, Staining, Marker, Injection, Comparison