wnt signaling agonist  (MedChemExpress)

Journal: Advanced Science

Article Title: Targeting the CRYAB/β‐Catenin Axis with an MMP‐9‐Responsive Hydrogel Disrupts CTC Clusters to Suppress Progression of Hepatocellular Carcinoma after Radiofrequency Ablation

doi: 10.1002/advs.77578

Figure Lengend Snippet: CRYAB stabilizes β‐catenin by suppressing ubiquitin‐dependent degradation to promote EMT and N2 polarization. (A) Molecular docking model showing the binding interface between CRYAB (red) and β‐catenin (blue). Hydrophobic interactions (left, residues LEU7, PRO62, VAL349, ARG6, ARG57) and hydrogen bonds (right, residues ARG342, GLN302, ASN387, ARG386, TRP383, LEU263) collectively stabilize the complex. (B) RMSD trajectories from 100‐ns molecular dynamics simulations, showing stable conformational dynamics for both CRYAB (∼0.35 nm) and β‐catenin (∼0.80 nm), confirming a robust and stable interaction interface. (C) Co‐IP of β‐catenin with CRYAB in control and heat‐stressed cells ( n = 3). (D) Ubiquitination assay (IP: β‐catenin; IB: Ub) showing reduced β‐catenin ubiquitination upon heat stress ( n = 3). (E) Western blot of β‐catenin in control and heat‐stressed HCC cells treated with MG132 (proteasome inhibitor) or vehicle ( n = 3). (F) Nuclear/cytoplasmic fractionation and western blot for CRYAB and β‐catenin in heat‐stressed HCC cells. Histone H3 serves as a nuclear marker ( n = 3). (G) Immunofluorescence image of nuclear CRYAB and β‐catenin in control and heat‐stressed HCC cells (n = 3). Scale bar: 50 µm. (H) Representative immunofluorescence images and quantification of TAN morphology cultured with si Cryab and BML‐284 treatment ( n = 3). (I) Representative immunofluorescence images and quantification of TAN morphology cultured with Cryab OE and XAV‐939 treatment ( n = 3). Scale bar: 10 µm. (J) FCM analysis and quantification of iNOS + TANs and CD206 + TANs cultured with si Cryab and BML‐284 treatment ( n = 3). (K) FCM analysis and quantification of iNOS + TANs and CD206 + TANs cultured with Cryab OE and XAV‐939 treatment ( n = 3). (L–O) qPCR ( n = 5) and ELISA ( n = 4) analyses of TGF‐β and MMP‐9 expression and secretion in the si Cryab +BML‐284 and Cryab OE +XAV‐939 rescue systems, respectively. (Data are means ± SD. Calculated by Student's t ‐test or One‐way ANOVA. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001).

Article Snippet: BML‐284 (purity ≥98%) was obtained from TargetMol (USA, 2095432‐75‐8).

Techniques: Ubiquitin Proteomics, Docking Assay, Binding Assay, Co-Immunoprecipitation Assay, Control, Western Blot, Fractionation, Marker, Immunofluorescence, Morphology, Cell Culture, Analysis, Enzyme-linked Immunosorbent Assay, Expressing, FACS

Journal: Diabetes & Metabolism Journal

Article Title: E2F5 Accelerates Vascular Smooth Muscle Cells Phenotype Switching in Diabetic Atherosclerosis through Activating Wnt/β-Catenin Pathway

doi: 10.4093/dmj.2024.0588

Figure Lengend Snippet: E2F transcription factor 5 (E2F5) knockdown repressed vascular smooth muscle cells (VSMCs) phenotype switching through inactivating Wnt/β-catenin pathway. (A) Western blot for detecting wnt1 and β-catenin expression in VSMCs in different groups. (B) 5-ethynyl-2´-deoxyuridine (EdU) assay was employed for assessing the proliferative ability of VSMCs (×200). (C) Flow cytometry was employed for testing VSMCs cell cycle. (D) Transwell assay was utilized for evaluating the migrative ability of VSMCs (×100). (E) Western blot for detecting α-smooth muscle actin (α-SMA), SM22, and osteopontin (OPN) expression in VSMCs. GAPDH, glyceraldehyde 3-phosphate dehydrogenase; HG, high glucose; ox-LDL, oxidized low-density lipoprotein; si-NC, negative control siRNA; BML-284, a Wnt pathway activator; DAPI, 4´,6-diamidino-2-phenylindole. a P <0.05, b P <0.05, c P <0.05.

Article Snippet: Meanwhile, VSMCs were treated with BML-284 (a Wnt pathway activator, Med-ChemExpress, Monmouth Junction, NJ, USA) for 48 hours.

Techniques: Knockdown, Western Blot, Expressing, EdU Assay, Flow Cytometry, Transwell Assay, Negative Control