polyclonal rabbit anti p21  (Cell Signaling Technology Inc)

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