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

Journal: bioRxiv

Article Title: p53Y220C-BET-bifunctionals (tPRIMEs) drive p53Y220C-mutant cancer cells into apoptosis

doi: 10.1101/2025.08.13.669398

Figure Lengend Snippet: p53Y220C-tPRIMEs markedly induce p53 target genes. A. Hierarchical cluster analysis of genes identified as significantly upregulated in RNA-sequencing by 24h treatment with tPRIME-2, tPRIME-3 or control ligands compared to DMSO. Pathway analysis of cluster of genes significantly upregulated by tPRIME-2/3 compared to control ligands. B. qPCR analysis of the p53 target genes CDKN1A/p21, MDM2, BBC3, ATF3, TP53I3 in NUGC3 cells treated with serial dilution of tPRIME-3, tPRIME-5 or control ligands. C. Western Blot analysis of p53-dependent and apoptotic proteins in NUGC3 cells treated for 24h with 1μM of tPRIME-5 or control ligands. D. LC-MS/MS analysis of proteomic changes across multiple p53Y220C-mutant cell lines treated for 24h with 1μM of tPRIME-5 compared to p53Y220C+BET-ligand combination.

Article Snippet: The following list of primary antibodies was used: anti-p53 (SantaCruz, #sc-126), anti-p21 (SantaCruz, #sc- 53870), anti-NQO1 (CST, #3187), anti-TP53I3 (Origene, #TA503656), anti-PUMA (CST, #12450), anti-cleaved Parp (CST, #5625), anti-Parp (CST, #9542), anti-MDM2 (CST, #86934), anti-BRD2 (CST, #5848), anti-BRD3 (CST, #94032), anti-BRD4 (CST, #13440), anti-GAPDH (CST, #97166), anti-β-actin (CST, #3700).

Techniques: RNA Sequencing, Control, Serial Dilution, Western Blot, Liquid Chromatography with Mass Spectroscopy, Mutagenesis

Journal: bioRxiv

Article Title: p53Y220C-BET-bifunctionals (tPRIMEs) drive p53Y220C-mutant cancer cells into apoptosis

doi: 10.1101/2025.08.13.669398

Figure Lengend Snippet: tPRIME-5 induces p53 target genes and tetramer formation at slower kinetics than p53Y220C-ligands. A. qPCR analysis of the p53 target genes CDKN1A, MDM2, BBC3, ATF3, TP53I3 in NUGC3 cells treated with 0.1 or 1μM of tPRIME-5 or p53Y220C-ligand over a time course of 2-24h. B. Western Blot analysis and quantification of p53 tetramer after DSG crosslinking in NUGC3 cells treated for 2-24h with 0.1μM, 1μM or 10μM p53Y220C-ligand or tPRIME-5.

Article Snippet: The following list of primary antibodies was used: anti-p53 (SantaCruz, #sc-126), anti-p21 (SantaCruz, #sc- 53870), anti-NQO1 (CST, #3187), anti-TP53I3 (Origene, #TA503656), anti-PUMA (CST, #12450), anti-cleaved Parp (CST, #5625), anti-Parp (CST, #9542), anti-MDM2 (CST, #86934), anti-BRD2 (CST, #5848), anti-BRD3 (CST, #94032), anti-BRD4 (CST, #13440), anti-GAPDH (CST, #97166), anti-β-actin (CST, #3700).

Techniques: Western Blot

Journal: bioRxiv

Article Title: p53Y220C-BET-bifunctionals (tPRIMEs) drive p53Y220C-mutant cancer cells into apoptosis

doi: 10.1101/2025.08.13.669398

Figure Lengend Snippet: tPRIME-5 induces p53 target genes in vivo and achieves tumor growth inhibition and regression in NUGC3 xenografts. A. Plasma pharmacokinetics and NUGC3 tumor pharmacodynamics measured post a single dose. Left y-axis shows plasma concentration of tPRIME-5 or p53Y220C-ligand measured 0.5h, 1h, 4h, 8h or 24h post dosing mice IV, IP or PO. Right y-axis shows gene expression changes of CDKN1A, BBC3 and TP53I3 in NUGC3 xenografts measured at 4h, 8h and 24h post a single dose. B. Schematic of 21-day efficacy study in NUGC3 xenograft model. C. NUGC3 tumor growth inhibition and body weight changes of individual animals across the ten treatment groups at the end of study.

Article Snippet: The following list of primary antibodies was used: anti-p53 (SantaCruz, #sc-126), anti-p21 (SantaCruz, #sc- 53870), anti-NQO1 (CST, #3187), anti-TP53I3 (Origene, #TA503656), anti-PUMA (CST, #12450), anti-cleaved Parp (CST, #5625), anti-Parp (CST, #9542), anti-MDM2 (CST, #86934), anti-BRD2 (CST, #5848), anti-BRD3 (CST, #94032), anti-BRD4 (CST, #13440), anti-GAPDH (CST, #97166), anti-β-actin (CST, #3700).

Techniques: In Vivo, Inhibition, Clinical Proteomics, Drug discovery, Concentration Assay, Gene Expression

Journal: bioRxiv

Article Title: p53Y220C-BET-bifunctionals (tPRIMEs) drive p53Y220C-mutant cancer cells into apoptosis

doi: 10.1101/2025.08.13.669398

Figure Lengend Snippet: tPRIME-5 induces marked expression of multiple p53 target genes in NUGC3 xenografts post a single dose. A. qPCR analysis of the p53 target genes CDKN1A, GDF15, MDM2, ATF3, BBC3, RRAD, NQO1, TP53I3 and ALDH1A3 in NUGC3 xenografts treated with a single dose of vehicle, 100mpk p53Y220C-ligand (PO), 30mpk BET-ligand (PO), 100mpk p53Y220C-ligand + 30mpk BET-ligand (PO), 100mpk tPRIME-5 (IV) or 100mpk tPRIME-5 (IP) and harvested 4h, 8h and 24h post dosing. B. Analysis of p53 dependent target proteins via Western Blot in NUGC3 xenograft samples harvested 4h, 8h and 24h post a single dose of treatment with vehicle, 100mpk p53Y220C-ligand (PO) or 100mpk tPRIME-5 (IV or IP).

Article Snippet: The following list of primary antibodies was used: anti-p53 (SantaCruz, #sc-126), anti-p21 (SantaCruz, #sc- 53870), anti-NQO1 (CST, #3187), anti-TP53I3 (Origene, #TA503656), anti-PUMA (CST, #12450), anti-cleaved Parp (CST, #5625), anti-Parp (CST, #9542), anti-MDM2 (CST, #86934), anti-BRD2 (CST, #5848), anti-BRD3 (CST, #94032), anti-BRD4 (CST, #13440), anti-GAPDH (CST, #97166), anti-β-actin (CST, #3700).

Techniques: Expressing, Western Blot

Journal: bioRxiv

Article Title: p53Y220C-BET-bifunctionals (tPRIMEs) drive p53Y220C-mutant cancer cells into apoptosis

doi: 10.1101/2025.08.13.669398

Figure Lengend Snippet: tPRIME-5 induces marked expression of multiple p53 target genes in NUGC3 xenografts harvested at the end of the efficacy study. A. qPCR analysis of the p53 target genes CDKN1A, GDF15, MDM2, ATF3, BBC3, RRAD, NQO1, TP53I3 and ALDH1A3 in NUGC3 xenografts treated for 21 days with vehicle, 100mpk p53Y220C-ligand (PO QD), 30mpk BET-ligand (PO QD), 100mpk p53Y220C-ligand + 30mpk BET-ligand (PO QD), 100mpk tPRIME-5 (IV BIW) or 100mpk tPRIME-5 (IP QD) and harvested 4h, 8h and 24h post last dose. B. Analysis of p53 dependent target proteins via Western Blot in NUGC3 xenograft samples harvested 4h, 8h and 24h post last treatment with vehicle, 100mpk p53Y220C-ligand (PO QD) or 100mpk tPRIME-5 (IV BIW or IP QD).

Article Snippet: The following list of primary antibodies was used: anti-p53 (SantaCruz, #sc-126), anti-p21 (SantaCruz, #sc- 53870), anti-NQO1 (CST, #3187), anti-TP53I3 (Origene, #TA503656), anti-PUMA (CST, #12450), anti-cleaved Parp (CST, #5625), anti-Parp (CST, #9542), anti-MDM2 (CST, #86934), anti-BRD2 (CST, #5848), anti-BRD3 (CST, #94032), anti-BRD4 (CST, #13440), anti-GAPDH (CST, #97166), anti-β-actin (CST, #3700).

Techniques: Expressing, Western Blot

Journal: Renal Failure

Article Title: Canagliflozin ameliorates high-salt-induced renal injury and premature aging in male Dahl salt-sensitive rats, with associated changes in SIRT6/HIF-1α signaling

doi: 10.1080/0886022X.2025.2546624

Figure Lengend Snippet: Representative images of senescence-associated β-galactosidase (SA-β-gal) staining and the expression of aging-related proteins in the kidneys of the three groups of rats. A. Renal SA-β-gal staining. Magnification, ×200; scale bar, 50 μm. B. Expression of the aging-related proteins P53 and P21 in the kidneys. C. SA-β-gal-positive area in rat kidneys ( n = 4). D. P53 protein expression. E. P21 protein expression. (D, E, n = 6). NSD: normal salt dahl salt-sensitive rats; HSD: high salt dahl salt-sensitive rats; HSD + CANA: high salt dahl salt-sensitive rats + canagliflozin (30 mg/kg/day). HSD vs. NSD: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. HSD vs. HSD + CANA: # p < 0.05, ## p < 0.01, ### p < 0.001, #### p < 0.0001.

Article Snippet: The antibodies used for rat tissues included rabbit monoclonal anti-SIRT6 (1:1000, Abcam, ab191385), rabbit polyclonal anti-p53 (1:800, Proteintech, 10442-1-AP), rabbit polyclonal anti-p21 (1:5000, Proteintech, 10355-1-AP), rabbit polyclonal anti-HIF-1α (1:100, affinity, AF1009), and rabbit monoclonal anti-αSMA (alpha smooth muscle, 1:1000, Zenbio, R380653) antibodies.

Techniques: Staining, Expressing

Journal: Renal Failure

Article Title: Canagliflozin ameliorates high-salt-induced renal injury and premature aging in male Dahl salt-sensitive rats, with associated changes in SIRT6/HIF-1α signaling

doi: 10.1080/0886022X.2025.2546624

Figure Lengend Snippet: Immunohistochemical results of the three groups of rats. A. Representative image of SIRT6 in the kidney. B. Representative image of HIF-1α in the kidney. C. Representative image of α-SMA in the kidney. D. Representative image of P53 in the kidney. E. Representative image of P21 in the kidney. Magnification, ×200; scale bar, 20 μm. NSD: normal salt dahl salt-sensitive rats; HSD: high salt dahl salt-sensitive rats; HSD + CANA: high salt dahl salt-sensitive rats + canagliflozin (30 mg/kg/day). HSD vs. NSD: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. HSD vs. HSD + CANA: # p < 0.05, ## p < 0.01, ### p < 0.001, #### p < 0.0001 ( n = 6).

Article Snippet: The antibodies used for rat tissues included rabbit monoclonal anti-SIRT6 (1:1000, Abcam, ab191385), rabbit polyclonal anti-p53 (1:800, Proteintech, 10442-1-AP), rabbit polyclonal anti-p21 (1:5000, Proteintech, 10355-1-AP), rabbit polyclonal anti-HIF-1α (1:100, affinity, AF1009), and rabbit monoclonal anti-αSMA (alpha smooth muscle, 1:1000, Zenbio, R380653) antibodies.

Techniques: Immunohistochemical staining

Journal: Frontiers in Pharmacology

Article Title: Exploring the potential anti-senescence effects of soybean-derived peptide Soymetide in mice hippocampal neurons via the Wnt/β-catenin pathway

doi: 10.3389/fphar.2025.1510337

Figure Lengend Snippet: Soymetide prevents doxorubicin-induced senescence in the hippocampus of mice . Hippocampal tissues were isolated from vehicle (V), doxorubicin (DOX), Soymetide (Soym)+DOX and senolytic combination (Seno)+DOX-treated mice. (A–F) Representative Western blots and corresponding densitometry (relative to (Rel.) vehicle) of p53 (A, D) , p21 (B, E) , and p16 (C, F) normalized with β-actin in the hippocampal tissues. Data represent means ± Standard Error (SE) of three mice/group. ***p < 0.001, **p < 0.01 and *p < 0.05 compared to V. ### p < 0.001, ## p < 0.01 and # p < 0.05 compared to DOX.

Article Snippet: Mouse monoclonal p53 (cat no. 2524), rabbit polyclonal p21 (cat no. 64016), phospho-LRP6 (cat no. 2568), β-catenin (cat no. 9562), rabbit monoclonal NeuN (cat no. 24307), LRP6 (cat no. 3395), Dvl2 (cat no. 3224), Axin1 (cat no. 2087), GSK-3β (cat no. 12456) and TCF3/TCF7L1 (cat no. 2883) antibodies were purchased from Cell Signaling Technology.

Techniques: Isolation, Western Blot

Journal: Frontiers in Pharmacology

Article Title: Exploring the potential anti-senescence effects of soybean-derived peptide Soymetide in mice hippocampal neurons via the Wnt/β-catenin pathway

doi: 10.3389/fphar.2025.1510337

Figure Lengend Snippet: The Soymetide-induced reduction in senescence markers and increase in NeuN levels are Wnt3a/β-catenin-dependent in the hippocampus of doxorubicin-treated mice . Hippocampal tissues and the whole brain were isolated from vehicle (V), doxorubicin (DOX), Soymetide (Soym)+DOX, Soym + DOX + rDkk1, and Soym + DOX + iCRT3-treated mice. (A–C, E) Representative Western blots and corresponding densitometry (relative to vehicle) of p53 (A) , p21 (B) , p16 (C) and NeuN (E) normalized with β-actin in the hippocampal tissues. (D) Representative photomicrographs and corresponding bar graph (relative to vehicle) of Senescence β-Galactosidase-stained hippocampus in brain sections. Scale bar: 100 µm. Data represent means ± SE of three mice/group. The quantification of the first three groups in (D) and (E) are based on the same data sets as in . ***p < 0.001 and **p < 0.01 compared to V. ### p < 0.001, ## p < 0.01 and # p < 0.05 compared to DOX. @@@ p < 0.001, @@ p < 0.01 and @ p < 0.05 compared to Soym + DOX.

Article Snippet: Mouse monoclonal p53 (cat no. 2524), rabbit polyclonal p21 (cat no. 64016), phospho-LRP6 (cat no. 2568), β-catenin (cat no. 9562), rabbit monoclonal NeuN (cat no. 24307), LRP6 (cat no. 3395), Dvl2 (cat no. 3224), Axin1 (cat no. 2087), GSK-3β (cat no. 12456) and TCF3/TCF7L1 (cat no. 2883) antibodies were purchased from Cell Signaling Technology.

Techniques: Isolation, Western Blot, Staining

Journal: Frontiers in Pharmacology

Article Title: Exploring the potential anti-senescence effects of soybean-derived peptide Soymetide in mice hippocampal neurons via the Wnt/β-catenin pathway

doi: 10.3389/fphar.2025.1510337

Figure Lengend Snippet: Schematic of the anti-senescent effect induced by Soymetide against doxorubicin treatment in hippocampal neurons and its positive impact on cognitive function . Soymetide prevents the reduction in Wnt3a and its related proteins (Fz, p-LRP-6, Dvl, and Axin1) caused by doxorubicin, and it counters the rise in GSK-3β and the reduction of β-catenin. This inhibits the increase in senescence markers such as p53, p21, p16, β- Galactosidase activity, and SASPs, as well as the decrease in neuronal survival and learning-memory functions. In summary, Soymetide exhibits anti-senescent effects, provides neuroprotection, and induces cognitive restoration by restoring the Wnt/β-catenin pathway.

Article Snippet: Mouse monoclonal p53 (cat no. 2524), rabbit polyclonal p21 (cat no. 64016), phospho-LRP6 (cat no. 2568), β-catenin (cat no. 9562), rabbit monoclonal NeuN (cat no. 24307), LRP6 (cat no. 3395), Dvl2 (cat no. 3224), Axin1 (cat no. 2087), GSK-3β (cat no. 12456) and TCF3/TCF7L1 (cat no. 2883) antibodies were purchased from Cell Signaling Technology.

Techniques: Activity Assay