Journal: British Journal of Cancer

Article Title: The predictive value of molecular markers (p53, EGFR, ATM, CHK2) in multimodally treated squamous cell carcinoma of the oesophagus

doi: 10.1038/sj.bjc.6604037

Figure Lengend Snippet: Antibodies used in this study

Article Snippet: p53, phosphorylated at Ser15 , 3A36 , United States Biological, Swampscott, MA, USA , 1 : 75 , Citrate, pH 6; heating 7 min.

Techniques:

Journal: Radiation Oncology (London, England)

Article Title: IL-6 signaling promotes DNA repair and prevents apoptosis in CD133+ stem-like cells of lung cancer after radiation

doi: 10.1186/s13014-015-0534-1

Figure Lengend Snippet: IL-6 knocked down CD133+ cells showed lower expression of DNA repair-associated molecules. a Higher expression of γ-H2AX indicating unrepaired DNA damage was found in IL-6 knocked down CD133+ cell cultures measured by IF staining. CD133+ cells of A549IL-6si/sc and H157IL-6si/sc pairs were irradiated with 6 Gy gamma rays and γ-H2AX IF staining was performed at 3 h after irradiation. b Higher DNA strand breaks were detected in CD133+ cells of IL-6si cells than those of sc cells. CD133+ and CD133- cells were obtained from A549IL-6si/sc and H157IL-6si/sc cell pairs, irradiated (6 Gy), and DNA strand breaks were analyzed at 0 and 30 min after radiation in Comet assay. The ratio of head diameter to comet length of 50 cells were measured in each sample analysis and used in quantitation shown on right. c Lower expression of DNA repair associated proteins, ATM, CHK2, p-ATM, and p-p53 were detected in CD133+ cells of IL-6 knocked down cell line than those of sc counterparts (IF staining). CD133+ and CD133- cells of A549IL-6si/sc and H157IL-6si/sc pairs were irradiated with 6 Gy gamma rays and IF staining was performed at 2 h after irradiation using antibodies of ATM, CHK2, p-ATM, and p-p53. d qPCR analysis. CD133+ cells of A549IL-6si/sc and H157IL-6si/sc pairs were irradiated with 6 Gy gamma rays and total RNAs were obtained for qPCR analyses at 6 h after irradiation. e ATM-luc assay. 293HEK and H1299 cells were transfected with ATM-luc and cultured with various concentrations of IL-6. At the end of 48 h of incubation, luciferase activities were measured

Article Snippet: Antibodies of ATM and CHK2 were obtained from Bethyl Laboratory (Montgomery, TX), phosphorylated ATM (Ser 1981), and phosphorylated p53 (Ser 20) were from Gene Tex (Irvine CA), and γ-H2AX antibody was purchased from Trevigen (Gaithersburg, MD).

Techniques: Expressing, Staining, Irradiation, Single Cell Gel Electrophoresis, Quantitation Assay, Transfection, Cell Culture, Incubation, Luciferase

Journal: International Journal of Molecular Sciences

Article Title: Evaluation of the Role of p53 Tumour Suppressor Posttranslational Modifications and TTC5 Cofactor in Lung Cancer

doi: 10.3390/ijms222413198

Figure Lengend Snippet: Example of the studied proteins staining in human lung cancer tissues. ( A , B ) positive staining of total p53 protein (p53T); ( C , D ) p53 phosphorylated at serine 46 (p53 S46); ( E , F ) TTC5 protein; and ( I , J ) SIRT1 at magnification 10× ( A , C , E , G , I ) and 40× ( B , D , F , H , J ). ( I , J ) is a control staining of lung cancer tissue with IgG antibody used as primary antibody. Scale bars indicate 250 µm for 10× and 62 µm for 40× magnification.

Article Snippet: The antibody for p53 (DO-7, 1:100 dilution) was purchased from DAKO (CA, USA), Phosphorylated p53 (Ser46 (1:100 dilution) was obtained from Abnova (Taipei, Taiwan), acetylated p53 (K382) (1:50 dilution) was from Abcam, Cambridge, UK.

Techniques: Staining, Control

Journal: International Journal of Molecular Sciences

Article Title: Evaluation of the Role of p53 Tumour Suppressor Posttranslational Modifications and TTC5 Cofactor in Lung Cancer

doi: 10.3390/ijms222413198

Figure Lengend Snippet: Association of total p53 (Tp53) with clinicopathological features.

Article Snippet: The antibody for p53 (DO-7, 1:100 dilution) was purchased from DAKO (CA, USA), Phosphorylated p53 (Ser46 (1:100 dilution) was obtained from Abnova (Taipei, Taiwan), acetylated p53 (K382) (1:50 dilution) was from Abcam, Cambridge, UK.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Evaluation of the Role of p53 Tumour Suppressor Posttranslational Modifications and TTC5 Cofactor in Lung Cancer

doi: 10.3390/ijms222413198

Figure Lengend Snippet: Association of acetylated p53 (p53) form (K382) with clinicopathological features.

Article Snippet: The antibody for p53 (DO-7, 1:100 dilution) was purchased from DAKO (CA, USA), Phosphorylated p53 (Ser46 (1:100 dilution) was obtained from Abnova (Taipei, Taiwan), acetylated p53 (K382) (1:50 dilution) was from Abcam, Cambridge, UK.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Evaluation of the Role of p53 Tumour Suppressor Posttranslational Modifications and TTC5 Cofactor in Lung Cancer

doi: 10.3390/ijms222413198

Figure Lengend Snippet: Association of the acetylated p53 (K382) subcellular localization with clinicopathological features.

Article Snippet: The antibody for p53 (DO-7, 1:100 dilution) was purchased from DAKO (CA, USA), Phosphorylated p53 (Ser46 (1:100 dilution) was obtained from Abnova (Taipei, Taiwan), acetylated p53 (K382) (1:50 dilution) was from Abcam, Cambridge, UK.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Evaluation of the Role of p53 Tumour Suppressor Posttranslational Modifications and TTC5 Cofactor in Lung Cancer

doi: 10.3390/ijms222413198

Figure Lengend Snippet: Association of the phosphorylated p53 (Ser-46) with clinicopathological features.

Article Snippet: The antibody for p53 (DO-7, 1:100 dilution) was purchased from DAKO (CA, USA), Phosphorylated p53 (Ser46 (1:100 dilution) was obtained from Abnova (Taipei, Taiwan), acetylated p53 (K382) (1:50 dilution) was from Abcam, Cambridge, UK.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Evaluation of the Role of p53 Tumour Suppressor Posttranslational Modifications and TTC5 Cofactor in Lung Cancer

doi: 10.3390/ijms222413198

Figure Lengend Snippet: Kaplan-Meier survival analysis. The subgroup KM curves are represented by different colours. The p -values are shown in the figure. The X axis shows the overall survival (OS) time in month. KM curves of the subgroups for total p53 ( A ), TTC5 ( B ), p53 K382 ( C ), p53 S46 ( D ), and SIRT1 ( E ) are shown.

Article Snippet: The antibody for p53 (DO-7, 1:100 dilution) was purchased from DAKO (CA, USA), Phosphorylated p53 (Ser46 (1:100 dilution) was obtained from Abnova (Taipei, Taiwan), acetylated p53 (K382) (1:50 dilution) was from Abcam, Cambridge, UK.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Evaluation of the Role of p53 Tumour Suppressor Posttranslational Modifications and TTC5 Cofactor in Lung Cancer

doi: 10.3390/ijms222413198

Figure Lengend Snippet: Univariate COX analysis. Status, expression (exp), and subcellular localization (loc) are shown.

Article Snippet: The antibody for p53 (DO-7, 1:100 dilution) was purchased from DAKO (CA, USA), Phosphorylated p53 (Ser46 (1:100 dilution) was obtained from Abnova (Taipei, Taiwan), acetylated p53 (K382) (1:50 dilution) was from Abcam, Cambridge, UK.

Techniques: Expressing

Journal: Journal of Medicinal Chemistry

Article Title: Fragment-based Differential Targeting of PPI Stabilizer Interfaces

doi: 10.1021/acs.jmedchem.9b01942

Figure Lengend Snippet: Identification of fragments binding to the interface of 14-3-3σ with p53. (A) Fragment AZ-001. (B) Crystal structure of AZ-001 (yellow sticks) in complex with 14-3-3σ (white surface) and p53pT387 (orange sticks), PDB 6S40. The final 2 F o – F c electron density maps are shown as blue mesh. (C) Detailed view of the binding pocket of AZ-001. The most prominent interaction is a salt-bridge (dotted black line) between the amidine of AZ-001 and carboxyl group of E14 of 14-3-3σ. (D) Fragment AZ-002. (E) Crystal structure of AZ-002 (yellow sticks) in complex with 14-3-3σ (white surface) and p53pT387 (orange sticks), PDB 6RWI. The final 2 F o – F c electron density maps are shown as blue mesh. (F) Detailed view of the binding pocket of AZ-002. Also with AZ-002, the most prominent interaction is a salt-bridge (dotted black line) between the amidine of AZ-002 and carboxyl group of E14 of 14-3-3σ.

Article Snippet: CTD Thr387 phosphorylated 32-mer peptide of p53 (AnaSpec) was immobilized onto a CMD200 M sensor chip (Xantec Bioanalytics) by using standard NHS/EDC amine-coupling chemistry.

Techniques: Binding Assay

Journal: Anatomy & Cell Biology

Article Title: Cyclosporin A aggravates hydrogen peroxide-induced cell death in kidney proximal tubule epithelial cells

doi: 10.5115/acb.18.192

Figure Lengend Snippet: Cyclosporin A (CsA) increases p53 activation and BH3 interacting-domain death agonist (BID) expression after H 2 O 2 injury in human kidney proximal tubule epithelial cells. HK-2 cells were treated with either 10 nM CsA or vehicle for 60 minutes and then exposed to 1 mM H 2 O 2 or control for 60 minutes (n=6 experiments per condition). (A) Phosphorylation level of p53 (p-p53, 53 kDa) and total expression level of p53 (t-p53, 53 kDa) were measured by western blot analysis. Antibody of β-actin (43 kDa) as a loading control was used for normalization. (B–D) Intensities of p-p53 and t-p53 protein bands were quantified using the AzureSpot software. (E) BID (22 kDa) expression was measured by western blot analysis. Antibody of β-actin (43 kDa) as a loading control was used for normalization. (F) Intensity of BID protein expression was quantified using the AzureSpot software. a) P <0.05 vs. control. b) P <0.05 vs. vehicle.

Article Snippet: These membranes were incubated with antibodies against phosphorylated p53 (p-p53; 1:2,500 dilution, catalog No. GTX70218, GeneTex, Irvine, TX, USA), total p53 (t-p53; 1:2,500 dilution, catalog No. sc-135630, Santa Cruz Biotechnology, Santa Cruz, CA, USA), BH3 interacting-domain death agonist (BID; 1:2,500 dilution, catalog No. sc-11423, Santa Cruz Biotechnology), phosphorylated-p38 (p-p38; 1:2,500 dilution, catalog No. 9211, Cell Signaling Technology, Beverly, MA, USA), total p38 (t-p38; 1:2,500 dilution, catalog No. 9212, Cell Signaling Technology), phosphorylated c-Jun N-terminal kinase (p-JNK; 1:2,500 dilution, catalog No. 9251, Cell Signaling Technology), total JNK (t-JNK; 1:5,000 dilution, catalog No. 9252, Cell Signaling Technology), phosphorylated extracellular signal-regulated kinase (p-ERK; 1:5,000 dilution, catalog No. 4370, Cell Signaling Technology), total ERK (t-ERK; 1:5,000 dilution, catalog No. sc-93, Santa Cruz Biotechnology), phosphorylated protein kinase B (p-AKT; 1:2,500 dilution, catalog No. 3787, Cell Signaling Technology), total AKT (t-AKT; 1:2,500 dilution, catalog No. sc-8312, Santa Cruz Biotechnology), manganese superoxide dismutase (MnSOD; 1:10,000 dilution, catalog No. sc-30080, Santa Cruz Biotechnology), copper/zinc superoxide dismutase (CuZnSOD; 1:10,000 dilution, catalog No. sc-11407, Santa Cruz Biotechnology), catalase (1:5,000 dilution, catalog No. sc-271803, Santa Cruz Biotechnology), 94 kDa glucoseregulated protein (GRP94; 1:5,000 dilution, catalog No. CSBPA00109A0Rb, CUSABIO, Wuhan, China), 78 kDa glucoseregulated protein (GRP78; 1:10,000 dilution, catalog No. CSB-PA010827YA01MO, CUSABIO), and β-actin (1:5,000 dilution, catalog No. sc-47778, Santa Cruz Biotechnology) overnight at 4°C, respectively.

Techniques: Activation Assay, Expressing, Control, Phospho-proteomics, Western Blot, Software