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: bioRxiv

Article Title: A fast 2C-induction method reveals a barrier role of SP2 for totipotency

doi: 10.64898/2026.07.23.740258

Figure Lengend Snippet: a , FACS analysis showing the number of MERVL-tdTomato positive cells from each induction condition. b , Upper panel: Bar plot showing the effect of each chemical on the induction of MERVL-tdTomato positive cells on the top. Data are mean ± s.d., n = 2 independent experiments. ** P <0.01, *** P <0.001; two-tailed unpaired Student’s t -test. Lower panel: Bar plots showing the expression of Nanog , Zscan4 , MERVL-int, and Dux from each condition. Data are mean ± s.d., n = 2 independent experiments. c , Left panel: FACS analysis showing the number of MERVL-tdTomato positive cells tested on C57 and 129 mESCs with 2C medium. Right panel: Bar plot showing the quantification of the corresponding FACS analysis. Data are mean ± s.d., n = 3 independent experiments. *** P < 0.001; two-tailed unpaired Student’s t -test. d , FACS analysis showing the number of MERVL-tdTomato positive cells tested on Dux KO mESCs in 2i and 2C medium. two independent experiments were shown.

Article Snippet: For 2C cell induction, mESCs were switched to 2C medium containing DMEM High-glucose, FBS (Gibco, 10099141), KnockOut SR (Gibco, 10828028), B27 (Gibco, 17504044), NEAA, 2-Mercaptoethanol, 100 nM AM580 (Selleck, S2933), 1 μM BML277 (TargetMOL, T2033), 1 μM JNK inhibitor VIII (MCE, HY-107598-1mg), 100 μM VPA (TargetMol, T7064), 5 μM EPZ5676 (Selleck, S7062), 5 mM LiCl (Sigma, 793620-500G), and 50 μg/mL Vitamin C (Sigma, 49752) for 36 h.

Techniques: Analysis, Two Tailed Test, Expressing

Journal: bioRxiv

Article Title: A fast 2C-induction method reveals a barrier role of SP2 for totipotency

doi: 10.64898/2026.07.23.740258

Figure Lengend Snippet: a , Schematic diagram for the induction of 2C-like cells from mESCs. Cells were first treated under 2i+LIF condition for at least 24h (stage 1), and then switch to 2C medium containing chemical molecules listed below (stage2). b , Merged cell images showing the morphological changes of mESCs during 2C medium at 0 h, 12 h, 24 h and 36 h with the overlapping of tdTomato signal on the top panel, and the FACS analysis showing the number of MERVL-tdTomato positive cells on the bottom panel. Scale bars = 250 μm. c , Expression changes of pluripotency genes ( Oct4 , Nanog , and Sox2 ) and totipotency genes ( Tcstv1 , Tcstv3 , and Zscan4 ) during 2C induction at 0 h, 12 h, 24 h and 36 h. Data are mean ± s.d., n = 2 independent experiments. d , Bar plot showing the percentage of MERVL-tdTomato positive cells from the dropout experiment. Data are mean ± s.d., n = 2 independent experiments. e , Bar plots showing the expression of Nanog , Zscan4 , MERVL-int, and Dux from each dropout condition. Data are mean ± s.d., n = 2 independent experiments.

Article Snippet: For 2C cell induction, mESCs were switched to 2C medium containing DMEM High-glucose, FBS (Gibco, 10099141), KnockOut SR (Gibco, 10828028), B27 (Gibco, 17504044), NEAA, 2-Mercaptoethanol, 100 nM AM580 (Selleck, S2933), 1 μM BML277 (TargetMOL, T2033), 1 μM JNK inhibitor VIII (MCE, HY-107598-1mg), 100 μM VPA (TargetMol, T7064), 5 μM EPZ5676 (Selleck, S7062), 5 mM LiCl (Sigma, 793620-500G), and 50 μg/mL Vitamin C (Sigma, 49752) for 36 h.

Techniques: Analysis, Expressing

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