Journal: Oncology Research

Article Title: MINDY1 Induces PD-L1 Deubiquitination to Promote Immune Escape in Hepatocellular Carcinoma by the Wnt/β-Catenin Pathway

doi: 10.32604/or.2025.067638

Figure Lengend Snippet: MINDY1 promotes PD-L1 deubiquitination through Wnt/β-catenin pathway. ( A–C ) MINDY1/Vector and sh-MINDY1/sh-NC were transfected, and Western blot analysis assessed that overexpression of MINDY1 elevated β-catenin and p-GSK3β(Tyr216)/GSK3β levels in HCC cells, and knockdown of MINDY1 did the opposite. ( D – F ) Western blot measured that SKL2001 (Wnt pathway activator) elevated the activating effect of overexpression of MINDY1 on the Wnt/β-catenin pathway, while pathway inhibitor LiCl decreased this phenomenon. (G) Western blot measured that SKL2001 increased PD-L1 protein expression, whereas LiCl led to PD-L1 downregulation. (H) Western blot examined that SKL2001 promoted the deubiquitination of PD-L1 in HCC cells, while LiCl suppressed its deubiquitination. n = 3. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. Vector/sh-NC; & p < 0.05, && p < 0.01, &&& p < 0.001 vs. MINDY1

Article Snippet: HCC cells transfected with MINDY1 were exposed to the Wnt inhibitor LiCl (20 mM, HY-Y0649, MedChemExpress, Monmouth Junction, NJ, USA) and incubated for 24 h, and this group was denoted as the MINDY1+LiCl group [ ].

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

Journal: Oncology Research

Article Title: MINDY1 Induces PD-L1 Deubiquitination to Promote Immune Escape in Hepatocellular Carcinoma by the Wnt/β-Catenin Pathway

doi: 10.32604/or.2025.067638

Figure Lengend Snippet: MINDY1 hinders CD8 + T cell activation through the Wnt/β-catenin pathway. ( A ) CFSE staining confirmed that co-culture with HCC cells transfected with MINDY1 resulted in diminished proliferation of CD8 + T cells, which was further diminished by SKL2001 treatment, whereas LiCl treatment did the opposite. ( B ) Flow cytometry indicated a rise in CD8 + T cell apoptosis after they were co-cultured with HCC cells overexpressing MINDY1, with an additional rise observed by SKL2001 treatment, while LiCl treatment reduced the apoptosis rate. ( C ) The cytotoxicity of CD8 + T cells was assessed through the LDH kit. ( D , E ) Flow cytometry confirmed that co-culturing with HCC cells overexpressing MINDY1 led to a decrease in Perforin + and TNF-α + T cells. After treatment with SKL2001, this decrease was further exacerbated, while LiCl increased activated CD8 + T cells. ( F , G ) ELISA was used to detect IFN-γ and IL-2 levels. ( H ) CCK-8 assay was employed to examine the survival rates of HCC cells. n = 3. * p < 0.05, *** p < 0.001 vs. Vector; & p < 0.05, && p < 0.01, &&& p < 0.001 vs. MINDY1

Article Snippet: HCC cells transfected with MINDY1 were exposed to the Wnt inhibitor LiCl (20 mM, HY-Y0649, MedChemExpress, Monmouth Junction, NJ, USA) and incubated for 24 h, and this group was denoted as the MINDY1+LiCl group [ ].

Techniques: Activation Assay, Staining, Co-Culture Assay, Transfection, Flow Cytometry, Cell Culture, Enzyme-linked Immunosorbent Assay, CCK-8 Assay, Plasmid Preparation

Journal: bioRxiv

Article Title: The structural mechanism of eukaryotic fluoride channel activation and inhibition by monovalent cations

doi: 10.64898/2026.04.21.719972

Figure Lengend Snippet: (A) Left, cryo-EM map of the FEX-CA/10E8v4 Fab complex. Right, atomic model of FEX-CA/10E8v4 Fab complex. For both panels, FEX-CA is shown in yellow, the 10E8v4 Fab heavy chain in salmon pink, and the light chain in green. Dashed lines indicate the approximate boundaries of the plasma membrane, with the cytoplasmic (in) and extracellular (out) sides labeled. (B) Structural comparison of FEX-CA in the presence of NaCl and LiCl analyzed using Resi-Ruler . Domain 1 of the Na⁺ and Li⁺ structures are superimposed and the magnitude of the per-residue C α distance differences is displayed as a color gradient (purple, Na + -FEX-CA; orange, Li + -FEX-CA). (C-D). Cross-sectional views of FEX-CA structures determined with Li⁺ (top) or Na⁺ (bottom). Cavities of the functional pore are shown from the extracellular view (C) or side (D), with cross-sectional regions at the same level, as indicated by yellow dashed lines. Protein surfaces of FEX-CA are colored by electrostatic potential as shown in the scale.

Article Snippet: Lysate was extracted with 2% n-Dodecyl-β-D-Maltoside (DDM) (Anatrace, Maumee, OH) for 2 hours at room temperature and protein purified using Strep-Tactin XT 4Flow resin (IBA Lifescience, Pittsburgh, PA) equilibrated with wash buffer (20 mM Tris-HCl, pH 8.0, 100 mM NaCl, 5 mM NaF, 1 mM DDM) followed by size-exclusion chromatography (SEC) (Superdex 200, Cytiva, Marlborough, MA) in SEC buffer (20 mM 4-(2-Hydroxyethyl)piperazine-1-ethanesulfonic acid (HEPES) pH 7.5, 150 mM LiCl, 5 mM NaF, and 0.5 mM DDM for cryo-EM or 1 mM DDM for functional studies.

Techniques: Cryo-EM Sample Prep, Clinical Proteomics, Membrane, Labeling, Comparison, Residue, Functional Assay

Journal: bioRxiv

Article Title: The structural mechanism of eukaryotic fluoride channel activation and inhibition by monovalent cations

doi: 10.64898/2026.04.21.719972

Figure Lengend Snippet: (A) Size exclusion chromatograms of MPER-tagged FEX-CA alone (gray dashed line) or with 10E8v4 Fab at a 1:1 molar ratio (blue trace) in the presence of 150 mM LiCl. Inset: Coomassie-stained SDS-PAGE of the peak fraction corresponding to the FEX-CA/10E8v4 Fab complex. (B) Representative cryo-EM micrograph and 2D averages of the FEX-CA/10E8v4 Fab complex in DDM micelles, with a 100 Å scale bar indicated. (C) Fourier shell correlation (FSC) curves as a function of resolution calculated between the half maps. (D) Local resolution map colored by local resolution estimate (B factor -137.8) using cryoSPARC. (E) Particle angular distribution plot.

Article Snippet: Lysate was extracted with 2% n-Dodecyl-β-D-Maltoside (DDM) (Anatrace, Maumee, OH) for 2 hours at room temperature and protein purified using Strep-Tactin XT 4Flow resin (IBA Lifescience, Pittsburgh, PA) equilibrated with wash buffer (20 mM Tris-HCl, pH 8.0, 100 mM NaCl, 5 mM NaF, 1 mM DDM) followed by size-exclusion chromatography (SEC) (Superdex 200, Cytiva, Marlborough, MA) in SEC buffer (20 mM 4-(2-Hydroxyethyl)piperazine-1-ethanesulfonic acid (HEPES) pH 7.5, 150 mM LiCl, 5 mM NaF, and 0.5 mM DDM for cryo-EM or 1 mM DDM for functional studies.

Techniques: Staining, SDS Page, Cryo-EM Sample Prep

Journal: RSC Advances

Article Title: Rational methodology of V–Al alloy-electrorefining protocols promotes high purity vanadium metal

doi: 10.1039/d5ra05411g

Figure Lengend Snippet: (a) CV curves in KCl–LiCl molten salt before (black dashed line) and after (red line) V–Al alloy dissolution for 6 h; (b) CV curves in KCl–LiCl molten salt after V–Al alloy dissolution for 6 h; (c) SWV of vanadium ions dissolved from V–Al alloy; (d) comparison of CV curves before and after adding F ions.

Article Snippet: In this study, a eutectic salt of KCl and LiCl in a molar ratio of 55 : 45 (KCl, purity 99.8%, LiCl, purity 99.8%, purchased from Macklin Biochemical Co., Ltd) was used as the electrolyte, with a total weight of 100 g. Before electrolysis, KCl and LiCl were mixed uniformly and vacuum-dried at 300 °C for 3 h to remove excess moisture, then heated to 500 °C and maintained at this temperature for 12 h for pre-electrolysis.

Techniques: Dissolution, Comparison

Journal: RSC Advances

Article Title: Rational methodology of V–Al alloy-electrorefining protocols promotes high purity vanadium metal

doi: 10.1039/d5ra05411g

Figure Lengend Snippet: (a) CV in KCl–LiCl after V–Al dissolution for 6 h at various scan rates; (b) the relationship between the current density and the square root of scan rate. Inset: relationship between peak potential and logarithm of scan rate; (c) CV in KCl–LiCl–KF after V–Al dissolution for 6 h at various scan rates. (d) The relationship between the current density and the square root of scan rate; (e) SWV in KCl–LiCl after V–Al dissolution for 6 h at various scan rates; (f) the relationship between the current density and the square root of frequency in KCl–LiCl; (g) SWV in KCl–LiCl–KF molten salt after V–Al dissolution for 6 h; (h) the relationship between the current density and the square root of frequency; MSD curve of V 3+ (i) before adding F ions; (j) after adding F ions; (k) the diffusion coefficient calculated by three methods before and after adding F ions.

Article Snippet: In this study, a eutectic salt of KCl and LiCl in a molar ratio of 55 : 45 (KCl, purity 99.8%, LiCl, purity 99.8%, purchased from Macklin Biochemical Co., Ltd) was used as the electrolyte, with a total weight of 100 g. Before electrolysis, KCl and LiCl were mixed uniformly and vacuum-dried at 300 °C for 3 h to remove excess moisture, then heated to 500 °C and maintained at this temperature for 12 h for pre-electrolysis.

Techniques: Dissolution, Diffusion-based Assay

Journal: RSC Advances

Article Title: Rational methodology of V–Al alloy-electrorefining protocols promotes high purity vanadium metal

doi: 10.1039/d5ra05411g

Figure Lengend Snippet: (a) Current curves in LiCl–KCl molten salt at −0.6 V; (b) plot of ( I / I m ) 2 and ( t / t m ) compared with theoretical curves at −0.6 V; (c) SEM images of the transient nucleation in the molten salt and the results of EDS analysis at point A; (d) current curves in LiCl–KCl molten salt at −0.9 V; (e) plot of ( I / I m ) 2 and ( t / t m ) compared with theoretical curves at −0.9 V; (f) SEM images of the progressive nucleation in the molten salt and the results of EDS analysis at point B.

Article Snippet: In this study, a eutectic salt of KCl and LiCl in a molar ratio of 55 : 45 (KCl, purity 99.8%, LiCl, purity 99.8%, purchased from Macklin Biochemical Co., Ltd) was used as the electrolyte, with a total weight of 100 g. Before electrolysis, KCl and LiCl were mixed uniformly and vacuum-dried at 300 °C for 3 h to remove excess moisture, then heated to 500 °C and maintained at this temperature for 12 h for pre-electrolysis.

Techniques: