Journal: Journal of Translational Medicine

Article Title: LDHC4 drives lung adenocarcinoma progression by inducing lactylation of RB1 at lysine 900 to disrupt the RB1–E2F1 complex

doi: 10.1186/s12967-026-08070-9

Figure Lengend Snippet: Influence of RB1–K900lac on the cell cycle pathway. ( A to C ) Cell cycle analysis by flow cytometry in A549 and PC-9 cell lines stably expressing RB1–WT or RB1–K900R via lentiviral vectors. ( D ) Representative immunofluorescence images showing the distribution of CDK1 in A549 and PC-9 cells. CDK1 protein was labeled with red fluorescent Cy3, and nuclei were counterstained with blue fluorescent DAPI. Images were acquired using a high-resolution confocal multiphoton microscopy system (NIKON AX RMP, Japan). ( E , F ) Western blot analysis of cell cycle-related CDK molecule expression. ( G , H ) Expression of cell cycle-related cyclin molecules. ( I , J ) Expression of P21 and Chk1 molecules. ( K ) Schematic diagram illustrating how LDHC4 promotes the cell cycle by inducing RB1 lactylation. ** p < 0.01, *** p < 0.001

Article Snippet: The following antibodies were used in this study: rabbit anti-human LDHC (subunit C) monoclonal antibody (mAb) (Proteintech Group, Inc., 1:1000), rabbit anti-human RB1 mAb (Proteintech Group, Inc., 1:2000), rabbit anti-human L-Lactyl Lysine mAb (PTM Bio, Inc., 1:1000), rabbit anti-human E2F1 mAb (APExBIO Technology, LLC, 1:500), rabbit anti-human Lamin B mAb (Beyotime Biotech, Inc., 1:1000), rabbit anti-human CDK1 mAb (Beyotime Biotech, Inc., 1:800), rabbit anti-human CDK2 mAb (Beyotime Biotech, Inc., 1:800), rabbit anti-human CDK4 mAb (Beyotime Biotech, Inc., 1:1000), rabbit anti-human CDK6 mAb (Beyotime Biotech, Inc., 1:1000), rabbit anti-human cyclin A2 mAb (Beyotime Biotech, Inc., 1:1000), rabbit anti-human cyclin B1 mAb (Beyotime Biotech, Inc., 1:1000), rabbit anti-human cyclin D1 mAb (Beyotime Biotech, Inc., 1:500), rabbit anti-human P21 mAb (Abmart Bio, Inc., 1:500), rabbit anti-human Chk1 mAb (Immunoway Bio, Inc., 1:2000), and rabbit anti-β-Actin monoclonal antibody (Beyotime Biotech, Inc., 1:1000). β-Actin and Lamin B served as loading controls, and protein band intensities were quantified using Image J software.

Techniques: Cell Cycle Assay, Flow Cytometry, Stable Transfection, Expressing, Immunofluorescence, Labeling, Microscopy, Western Blot

Journal: Scientific Reports

Article Title: Exploring the dual role of extracellular vesicles in coagulation and immune modulation in glioblastoma

doi: 10.1038/s41598-026-42867-4

Figure Lengend Snippet: Tumor-derived EVs reduce migration, induce senescence, and a M1-like phenotype in microglia. ( A ) Representative fluorescence microscopy images showing time-dependent uptake of EVs derived from different GBM cell lines by microglia. Scale bar = 20 μm ( B ) Quantitative analysis of EVs uptake confirms time- and cell line–dependent differences in internalization. n = 3, mean ± SD, Two-way ANOVA (Tukey’s multiple comparison test), ** p < 0.01, **** p < 0.0001. ( C ) Transwell migration assay reveals significantly reduced migratory capacity in microglia following EVs exposure. n = 3, mean ± SD, One-way ANOVA (Tukey’s multiple comparison test), ** p < 0.01. ( D , E ) Immunofluorescence staining and quantification show increased expression of senescence-associated markers p21, p16, and p53 in EVs-exposed microglia compared to untreated controls. Scale bar = 20 μm ( F , G ) Immunofluorescence and corresponding quantification indicate no significant changes in connexin43 or the proliferation marker Ki67, suggesting that EVs-induced senescence is not accompanied by altered proliferation or gap junctional communication. Scale bar = 20 μm ( H , I ) Immunofluorescence and corresponding quantification for iNOS demonstrate increased expression in microglia treated with GBM-derived EVs, indicating induction of an M1-like pro-inflammatory phenotype. LPS-treated microglia serve as a positive control. n = 3, mean ± SD, One-way ANOVA (Tukey’s multiple comparison test), * p < 0.05, *** p < 0.001. Scale bar = 20 μm.

Article Snippet: Senescence was evaluated using Alexa Fluor ® 594 p53 Ab (clone: DO-1, stock: 0.5 mg/ml, BioLegend, 1:250), Alexa Fluor ® 488 anti-human p21 Waf1/Cip1 Ab (clone: 12D1stock: 50 μg/ml, Cell Signaling Technology, 1:300), and Alexa Fluor ® 546 anti-human p16 Ab (clone: F-12, stock: 200 μg/ml, Santa Cruz Biotechnology, Dallas, Texas, United States, 1:50).

Techniques: Derivative Assay, Migration, Fluorescence, Microscopy, Comparison, Transwell Migration Assay, Immunofluorescence, Staining, Expressing, Marker, Positive Control

Journal: Antioxidants

Article Title: Exploring the Anticancer Potential of the Multistrain Probiotic Formulation OxxySlab in Bladder Cancer Cell Lines

doi: 10.3390/antiox14111282

Figure Lengend Snippet: Senescence induction in BC cells following OxxySlab lysate treatment. Cell lines were treated with OxxySlab lysate (75–150 μg/mL) for 48 h. Senescent cells were identified by β-galactosidase staining (β-gal). Representative phase-contrast images of T24 ( A ), 5637 ( E ) and SV-HUC-1 ( I ) β-gal-positive cells (blue staining) are shown. Quantification of senescence was performed by measuring the Mean Gray Value of blue staining intensity using ImageJ software, with data expressed as Inverted Mean Gray Value (255 − MGV), which directly reflects β-gal activity. Data are expressed as mean ± SEM of two independent experiments in triplicate. Western blot analysis of the senescence markers p21, p53 and p16 was performed in untreated (CNTR) and OxxySlab -treated T24 ( B – D ), 5637 ( F – H ), and SV-HUC1 ( J – L ) cells. Protein levels were quantified by densitometry, normalized to GAPDH, and expressed as fold change relative to CNTR of three independent experiments (mean ± SEM). Statistical significance was assessed by one-way ANOVA followed by Tukey post hoc test (* p < 0.05, ** p < 0.01).

Article Snippet: Following incubation with 5% non-fat dry milk in Tris-buffered saline for 1 h at room temperature, the membranes were incubated overnight at 4 °C with primary antibodies: goat anti-human vimentin polyclonal antibody (Chemicon International, Temecula, CA, USA; dilution 1:100), mouse anti-human E-cadherin monoclonal antibody (Cell Signaling Technology; dilution 1:1000), rabbit anti-human β-catenin monoclonal antibody (Cell Signaling Technology; dilution 1:1000), rabbit anti-human p21 polyclonal antibody (Santa Cruz Biotechnology, Dallas, TX, USA dilution 1:1000); rabbit anti-human phospho-Nrf2 monoclonal antibody (S40) (1:2000, Abcam, Cambridge, UK); rabbit anti-human p16 polyclonal antibody (Santa Cruz Biotechnology, dilution 1:200); mouse anti-human p53 monoclonal antibody (Santa Cruz Biotechnology, dilution 1:1000); mouse anti-human GAPDH monoclonal antibody (Immunological Sciences, Rome, Italy; dilution 1:1000).

Techniques: Staining, Software, Activity Assay, Western Blot