Journal: Oncology letters

Article Title: Expression of USP22 and the chromosomal passenger complex is an indicator of malignant progression in oral squamous cell carcinoma.

doi: 10.3892/ol.2018.9837

Figure Lengend Snippet: Figure 2. The effects of USP22 knockdown in OSCC cells. (A) USP22 siRNA were transfected into Ca9‑22 cells. At 48 h following transfection, cells were collected and the expression of Survivin, Aurora‑B, p21 and cyclin B was examined by western blot analysis. β‑actin was used as a control. (B) Cell growth of siRNA treated Ca9‑22 cells. At 48 h following USP22 siRNA treatment, 5,000 cells were plated on 24‑well plates. At 24 h, the cell number was counted as 0 day. The cell number was subsequently counted at days 2, 4, and 6. **P<0.01. USP22, ubiquitin‑specific protease 22; OSCC, oral squamous cell carcinoma; siRNA, small interfering RNA; p21, cyclin‑dependent kinase inhibitor 1A.

Article Snippet: The primary antibodies were USP22 (1:1,000; Abcam; cat. no. ab71732), Survivin (cat. no. NB500-201; Novus Biologicals; 1:2,000), Aurora-B (cat. no AMI-1; 1:000; Transduction Laboratories), Cyclin B (cat. no. 610219), p21 (cat. no. clone 70) (1:1,000; Transduction Laboratories) and β-actin (1:2,000, OriGene Technologies, Inc.; cat. no TA-09). siRNA transfection.

Techniques: Knockdown, Transfection, Expressing, Western Blot, Control, Small Interfering RNA

Journal: PLoS ONE

Article Title: The Role of Dlc1 Isoform 2 in K-Ras2 G12D Induced Thymic Cancer

doi: 10.1371/journal.pone.0040302

Figure Lengend Snippet: A(i) p21 waf1 protein in T-cell lymphoma and carcinoma cell lines, (ii) Plot showing the relative intensity of p21 waf1 protein in T-cell lymphoma and carcinoma cell lines relative to actin. Means and standard error of mean were determined from at least five independent experiments. (*p<0.05, **P<0.01, ***P<0.001, ****P<.0001). (B) Measurement of active RhoGTP in cell lines . (i) RhoGTP pull down assay showing constitutive and LPA induced levels of active RhoGTP in T-cell lymphoma and thymic epithelial carcinoma cell lines. (ii) Plot showing the RhoGTP to total Rho protein ratio by scanning the relative intensity of the protein bands in non-treated and LPA induced in TEC, TL, MEF cell lines. Means and standard error of mean determined from four independent experiments. (*p<0.05, **P<0.01, ***P<0.001 and ****P<.0001 by Student t test and two way ANOVA test.

Article Snippet: The western blots were hybridized with Dlc1 antibody (Sc32931, Santa Cruz Biotechnology, CA, USA) and p21 waf1 antibody (Acris Antibodies Gbmh, AP06263PU-N) and visualized.

Techniques: Pull Down Assay

Journal: Cell Death & Disease

Article Title: The embryonic transcription factor Brachyury blocks cell cycle progression and mediates tumor resistance to conventional antitumor therapies

doi: 10.1038/cddis.2013.208

Figure Lengend Snippet: Brachyury modulates expression of cell cycle regulatory proteins. Western blot analysis of Brachyury and the cell cycle regulatory proteins Rb, cyclin D1 and p21 was performed using ( a ) the A549 tumor cell pair at indicated times after release from cell cycle arrest by serum starvation or ( b ) asynchronous cultures of H460 control versus Brachyury shRNA-1 and -2 cells. ( c ) Two tumor cell lines derived from single-cell cloning of H460 cells were analyzed for expression of Brachyury in relation to Rb and p21. ( d ) H460 cells transfected with p21 expression vector or pCMV control were analyzed for p21 and Rb expression by western blot and ( e ) growth kinetics over a 5-day period. ( f ) Indicated cells were treated with cytotoxic therapies and assayed for survival in comparison with untreated cells. ( g ) The H460 cell pair transfected with a pool of nonspecific control siRNA or p21-specific siRNAs was treated with γ -radiation (1 Gy) and evaluated for cell death by using CellTiter-Glo (Promega). (** P <0.01; *** P <0.001)

Article Snippet: The full-length human Brachyury construct (pCMV-Neo-Brachyury, accession number NM_003181.2), a p21 expression vector (accession number NM_078467.1) and empty-vector control (pCMV-Neo) were purchased from Origene Technologies Inc. (Rockville, MD, USA).

Techniques: Expressing, Western Blot, shRNA, Derivative Assay, Clone Assay, Transfection, Plasmid Preparation

Journal: Cell Death & Disease

Article Title: The embryonic transcription factor Brachyury blocks cell cycle progression and mediates tumor resistance to conventional antitumor therapies

doi: 10.1038/cddis.2013.208

Figure Lengend Snippet: Brachyury drives repression of p21. ( a ) A ChIP assay was employed utilizing control IgG versus anti-Brachyury antibody with template DNA obtained from indicated cells. Shown at the top is the T-box palindromic consensus element juxtaposed with the half-binding site (shaded) located in the human p21 promoter (wild type), and a mutated p21 promoter containing point mutations in the T-box-binding site (denoted by arrowheads). Depicted below is the gel electrophoresis of PCR products amplified from the indicated immunoprecipitated DNA template sources using p21 promoter-specific primers. M corresponds to the DNA ladder. ( b ) Luciferase reporter assay was used to analyze H460 cells utilizing a wild-type p21 promoter driving the expression of the luciferase gene. ( c ) Luciferase reporter assay with H460 control.shRNA cells utilizing a wild-type versus mutated human p21 promoter driving the expression of the luciferase gene. Values shown represent the average of two technical replicates corrected for background±S.E.M. (* P <0.05). ( d ) Synchronized A549 pCMV and pBrachyury cells or asynchronous H460 control.shRNA and H460 Brachyury.shRNA2 were treated with indicated doses of radiation; cells collected after 24 h were analyzed for p21 induction by western blot. Intensity of p21 detection was quantitated and compared against β -actin or GAPDH (bottom panels)

Article Snippet: The full-length human Brachyury construct (pCMV-Neo-Brachyury, accession number NM_003181.2), a p21 expression vector (accession number NM_078467.1) and empty-vector control (pCMV-Neo) were purchased from Origene Technologies Inc. (Rockville, MD, USA).

Techniques: Binding Assay, Nucleic Acid Electrophoresis, Amplification, Immunoprecipitation, Luciferase, Reporter Assay, Expressing, shRNA, Western Blot

Journal: Molecular Medicine Reports

Article Title: Prognostic and predictive roles of microRNA-411 and its target STK17A in evaluating radiotherapy efficacy and their effects on cell migration and invasion via the p53 signaling pathway in cervical cancer

doi: 10.3892/mmr.2019.10826

Figure Lengend Snippet: Reverse transcription-quantitative polymerase chain reaction primer sequences.

Article Snippet: The membranes were incubated with phosphorylated (p)-STK17A antibody (cat. no. 14433-1-AP; Proteintech, Wuhan, China; 1:1,000), p-p21 WAF1 antibody (cat. no. AP01654PU-N; Origene; 1:10,000), p-p53 antibody (cat. no. MABE518; Merck KGaA; 1:1,000), TAp63 antibody (cat. no. TA311397; Origene; 1:1,000), and GAPDH antibody (cat. no. 10494-1-AP; Proteintech; 1:2,000) at 4°C overnight.

Techniques: Polymerase Chain Reaction, Sequencing

Journal: Molecular Medicine Reports

Article Title: Prognostic and predictive roles of microRNA-411 and its target STK17A in evaluating radiotherapy efficacy and their effects on cell migration and invasion via the p53 signaling pathway in cervical cancer

doi: 10.3892/mmr.2019.10826

Figure Lengend Snippet: miR-411 activates the p53 signaling pathway and negatively regulates STK17A in cervical cancer cells. (A) Determination by reverse transcription-quantitative polymerase chain reaction analysis demonstrated that ectopic expression of miR-411 and siRNA-mediated knockdown of STK17A decreased the mRNA expression of STK17A, but increased the mRNA expression of p53, p21 WAF1 and TAp63; miR-411 inhibitor increased the mRNA expression of STK17A, but decreased the mRNA expression of p53, p21 WAF1 and TAp63. (B) Determination by western blot analysis and (C) quantification demonstrated that ectopic expression of miR-411 and siRNA-mediated knockdown of STK17A decreased the protein expression of STK17A, but increased the protein expression of p53, p21 WAF1 and TAp63; miR-411 inhibitor increased the protein expression of STK17A, but decreased the protein expression of p53, p21 WAF1 and TAp63. *P<0.05, vs. NC group; # P<0.05, vs. miR-411 inhibitor + siRNA-STK17A group. The experiment was repeated three times and data were compared by one-way analysis of variance and analyzed by Tukey's post hoc test. NC, negative control; miR-411, microRNA-411; STK17A, serine/threonine kinase 17a; siRNA, small interfering RNA.

Article Snippet: The membranes were incubated with phosphorylated (p)-STK17A antibody (cat. no. 14433-1-AP; Proteintech, Wuhan, China; 1:1,000), p-p21 WAF1 antibody (cat. no. AP01654PU-N; Origene; 1:10,000), p-p53 antibody (cat. no. MABE518; Merck KGaA; 1:1,000), TAp63 antibody (cat. no. TA311397; Origene; 1:1,000), and GAPDH antibody (cat. no. 10494-1-AP; Proteintech; 1:2,000) at 4°C overnight.

Techniques: Reverse Transcription, Real-time Polymerase Chain Reaction, Expressing, Knockdown, Western Blot, Negative Control, Small Interfering RNA

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: Molecular medicine reports

Article Title: Growth inhibition effects of ent-11α-hydroxy-15-oxo-kaur-16-en-19-oic-acid on colorectal carcinoma cells and colon carcinoma-bearing mice.

doi: 10.3892/mmr.2016.4950

Figure Lengend Snippet: Figure 6. Effect of 5F treatment on the cell cycle, apoptosis and growth‑asso ciated proteins. Cells were treated with 20 µg/ml 5F for 24 h. (A) Western blots and (B) the relative density of p21, Bcl‑2, Bax, NF‑κB p65, IκB, survivin and β‑actin. **P<0.01 vs. the control. 5F, ent‑11α‑hydroxy‑15‑oxo ‑kaur‑16‑en‑19‑oic‑acid; p21, cyclin‑dependent kinase inhibitor 1A; Bcl‑2, B‑cell CLL/lymphoma 2; Bax, Bcl‑2‑associated X protein; NFκB, nuclear factor‑κ‑light‑chain enhancer of activated B cells; IκBα, nuclear factor of κ light polypeptide gene enhancer in B‑cells inhibitor, α.

Article Snippet: Immunoblots of 50 μg total protein were probed with primary antibodies overnight as follows: Rabbit anti-human polyclonal cyclin-dependent kinase inhibitor 1A (p21, Cip1) (p21; cat. no. BA0272; dilution, 1:200; Boster Bio-Engineering Ltd., Wuhan, China), rabbit anti-human polyclonal B-cell lymphoma 2 (Bcl-2; cat. no. ZS-492; dilution, 1:300; Zhongshan Golden Bridge Biotechnology, Co., Ltd., Beijing, China), mouse anti-mouse monoclonal Bcl-2-associated X protein (Bax; cat. no. ZS-7480; dilution, 1:300; Zhongshan Golden Bridge Biotechnology, Co., Ltd.), rabbit anti-human polyclonal nuclear factor κ-light-chain enhancer of activated B cells (NF-κB) p65 phosphorylated (p)-Ser536 (cat. no. 11014; dilution, 1:500; Signalway Antibody LLC., College Park, MD, USA), rabbit anti-human polyclonal nuclear factor of κ light polypeptide gene enhancer in B-cells inhibitor α (IκBα), rabbit anti-human polyclonal survivin (cat. no. sc-10811; dilution, 1:300; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) and β-actin (cat. no. sc-130656; dilution, 1:500; Santa Cruz Biotechnology, Inc.).

Techniques: Western Blot, Control