Journal: Scientific Reports

Article Title: Identification of cyclins A1, E1 and vimentin as downstream targets of heme oxygenase-1 in vascular endothelial growth factor-mediated angiogenesis

doi: 10.1038/srep29417

Figure Lengend Snippet: ( A ) Control siRNA (siCTL)-transfected or pooled siHO-1-transfected HUVEC were left untreated or treated with 25 ng/ml VEGF for 48 h prior to propidium iodide staining and flow-cytometric analysis of DNA distribution. Data is presented as mean ± SEM (n = 3 experiments), * p < 0.05 vs siCTL, δ p < 0.05 vs siCTL + VEGF. ( B ) HUVEC were exposed to geneFECTOR alone (GF) or transfected with control siRNA (CTL) or pooled HO-1 siRNA (HO-1). Cells were cultured in the presence or absence of VEGF for 24 h prior to mRNA quantification by qRT-PCR of ( B ) cyclin A1, ( C ) cyclin E1, ( D ) cyclin-dependent kinase 2 (cdk2) or ( E ) p27. Data are presented as mean ± SEM (n = 3 experiments), * p < 0.05, ** p < 0.01. δδ p < 0.01 vs untreated siCTL-transfected cells.

Article Snippet: Antibodies against Bcl-2, cyclin A1, proliferating cell nuclear antigen (PCNA) and retinoblastoma (Rb) protein were purchased from Santa Cruz Biotechnology (California, USA).

Techniques: Control, Transfection, Staining, Cell Culture, Quantitative RT-PCR

Journal: Scientific Reports

Article Title: Identification of cyclins A1, E1 and vimentin as downstream targets of heme oxygenase-1 in vascular endothelial growth factor-mediated angiogenesis

doi: 10.1038/srep29417

Figure Lengend Snippet: HUVEC were transfected with CTLAF 488 or HO-1 siRNA (seq2AF 488 ). Monolayers were scratched and treated with vehicle alone or VEGF (25 ng/ml) for 16 h ( A ) Migration was quantified by live cell imaging using ImageJ. EC were then harvested, lysed and immunoblotted for: ( B ) cyclin A1 and ( C ) retinoblastoma protein (pRb) and phospho-pRb pT821 . The histograms show corresponding densitometry data corrected for the GAPDH loading control and with phospho-pRb expressed relative to total pRb. Data are presented as mean ± SEM (n = 3 experiments), * p < 0.05, ** p < 0.01; *** p < 0.001.

Article Snippet: Antibodies against Bcl-2, cyclin A1, proliferating cell nuclear antigen (PCNA) and retinoblastoma (Rb) protein were purchased from Santa Cruz Biotechnology (California, USA).

Techniques: Transfection, Migration, Live Cell Imaging, Control

Journal: Scientific Reports

Article Title: Identification of cyclins A1, E1 and vimentin as downstream targets of heme oxygenase-1 in vascular endothelial growth factor-mediated angiogenesis

doi: 10.1038/srep29417

Figure Lengend Snippet: VEGF treatment of endothelial cells activates HO-1-dependent migration, proliferation and angiogenesis in vitro and in vivo . Microarray and proteomic analyses revealed cyclin A1, cyclin E1 and vimentin as downstream targets of HO-1. VEGF induced an HO-1-dependent increase in cdk2 kinase and calpain activity. These changes increased cell cycle progression and vimentin cleavage respectively. The role of HO-1 was established using specific siRNAs and HO-1 agonists. Similarly the importance of both cyclin A1 and vimentin in pro-angiogenic effects of VEGF and HO-1 activity were demonstrated by cyclin A1 and vimentin gene silencing.

Article Snippet: Antibodies against Bcl-2, cyclin A1, proliferating cell nuclear antigen (PCNA) and retinoblastoma (Rb) protein were purchased from Santa Cruz Biotechnology (California, USA).

Techniques: Migration, In Vitro, In Vivo, Microarray, Activity Assay

Journal: Nucleic Acids Research

Article Title: Dissecting the role of p53 phosphorylation in homologous recombination provides new clues for gain-of-function mutants

doi: 10.1093/nar/gkn503

Figure Lengend Snippet: Influence of cdk inhibition on HR regulation by cyclin A1 and mutant p53. ( A ) DSB-induced HR in KMV(HR/3′) cells treated with DMSO (−) or 80 μM olomoucine (+). Recombination frequencies in controls −p53/−cyclin A1/−olomoucine were set to 100% (absolute mean value: 1.1 × 10 −3 ). Columns, mean ( n = 12–18); bars, SEM. ( B ) Immunoblot analysis for p53pSer315 and p53. ( C ) Cell-cycle analysis. Columns, mean ( n = 2); bars, SD.

Article Snippet: For immunodetection of rat p53, we used rabbit serum NCL-p53-CM5p from Novocastra, for RAD51 H-92, for cyclin A1 H-230 from Santa Cruz Biotechnology, Santa Cruz, CA, USA and for cdk2 07-631 from Upstate.

Techniques: Inhibition, Mutagenesis, Western Blot, Cell Cycle Assay

Journal: Nucleic Acids Research

Article Title: Dissecting the role of p53 phosphorylation in homologous recombination provides new clues for gain-of-function mutants

doi: 10.1093/nar/gkn503

Figure Lengend Snippet: Effect of cyclin A1 and A2 on homologous DSB repair in cells differing in their p53 and topo I status. ( A ) DSB-induced HR was determined in KMV(HR/3′) cells cotransfected with pCMV-I-SceI, and pCMV-Wtp53, pCMV-p53(315A), pCMV-p53(273H), pCMV-p53(248W) or empty vector (control), and pcDNA3-cyclin A1 (A1), pCMV-cyclin A2 (A2) or empty vector (−). Columns, mean ( n = 12–27); bars, SEM. ( B ) DSB-induced HR in cells overexpressing cyclin A1, cotransfected with pSUPER ( − ) or pSUPER-TopoI (+). Columns, mean ( n = 12–18); bars, SEM. ( C ) DSB-induced HR in cells overexpressing cyclin A2, cotransfected with pSUPER (−) or pSUPER-TopoI (+). Columns, mean ( n = 6); bars, SEM. Recombination frequencies in controls were set to 100% (absolute mean value in (A) −p53/−cyclins: 2.1 × 10 −3 ; in (B) −p53/+cyclin A1/−pSUPER-TopoI: 2.2 × 10 −3 ; in (C) −p53/+cyclin A2/−pSUPER-TopoI: 2.8 × 10 −3 ). ( D ) pSUPER-TopoI-mediated downregulation of topo I protein compared to p53 expression. ( E ) Western blotting of cyclin A1 and cyclin A2.

Article Snippet: For immunodetection of rat p53, we used rabbit serum NCL-p53-CM5p from Novocastra, for RAD51 H-92, for cyclin A1 H-230 from Santa Cruz Biotechnology, Santa Cruz, CA, USA and for cdk2 07-631 from Upstate.

Techniques: Plasmid Preparation, Control, Expressing, Western Blot

Journal: Nucleic Acids Research

Article Title: Dissecting the role of p53 phosphorylation in homologous recombination provides new clues for gain-of-function mutants

doi: 10.1093/nar/gkn503

Figure Lengend Snippet: Regulation of HR by cyclin A1 and mutant p53 after knockdown of cdk2. ( A ) DSB-induced HR in KMV(HR/3′) cells cotransfected with empty vector or pKD-cdk2-v5 (pKD-cdk2) for cdk2-specific RNA interference. Recombination frequencies in controls −p53/−cyclin A1/−pKD-cdk2 were set to 100% (absolute mean value: 1.2 × 10 −3 ). Columns, mean ( n = 6); bars, SEM. ( B ) Western blot analysis of cdk2.

Article Snippet: For immunodetection of rat p53, we used rabbit serum NCL-p53-CM5p from Novocastra, for RAD51 H-92, for cyclin A1 H-230 from Santa Cruz Biotechnology, Santa Cruz, CA, USA and for cdk2 07-631 from Upstate.

Techniques: Mutagenesis, Knockdown, Plasmid Preparation, Western Blot

Journal: Nucleic Acids Research

Article Title: Dissecting the role of p53 phosphorylation in homologous recombination provides new clues for gain-of-function mutants

doi: 10.1093/nar/gkn503

Figure Lengend Snippet: Coimmunoprecipitation of topo I and p53 is enhanced by cyclin A1. KMV(HR/3′) cells were electroporated with pCMV-I-SceI plus empty expression vector (control), pCMV-p53(248W) or pCMV-Wtp53 plus empty vector or pcDNA3-cyclin A1, and treated with DMSO (−) or 80 μM olomoucine, exactly as described in . Twenty-four hours later, whole-cell extracts were prepared without [p53(248W)] or with (Wtp53) cross-linking of chromatin complexes in vivo and topo I immunoprecipitated with Scl-70 serum followed by western blotting for p53 or topo I. As a negative control for nonspecific topo I interactions, the extract was immunoprecipitated with anti-GAPDH antibodies (IgG). Nonspecific signals in the western blot from the heavy chain with a molecular mass similar to p53 were excluded by the empty vector controls.

Article Snippet: For immunodetection of rat p53, we used rabbit serum NCL-p53-CM5p from Novocastra, for RAD51 H-92, for cyclin A1 H-230 from Santa Cruz Biotechnology, Santa Cruz, CA, USA and for cdk2 07-631 from Upstate.

Techniques: Expressing, Plasmid Preparation, Control, In Vivo, Immunoprecipitation, Western Blot, Negative Control

Journal: Nucleic Acids Research

Article Title: Dissecting the role of p53 phosphorylation in homologous recombination provides new clues for gain-of-function mutants

doi: 10.1093/nar/gkn503

Figure Lengend Snippet: Proposed model of a cyclin A1-induced mechanism leading to genomic instability by oncogenic mutant p53.

Article Snippet: For immunodetection of rat p53, we used rabbit serum NCL-p53-CM5p from Novocastra, for RAD51 H-92, for cyclin A1 H-230 from Santa Cruz Biotechnology, Santa Cruz, CA, USA and for cdk2 07-631 from Upstate.

Techniques: Mutagenesis

Journal: Oncotarget

Article Title: Metformin mediates resensitivity to 5-fluorouracil in hepatocellular carcinoma via the suppression of YAP

doi: 10.18632/oncotarget.10079

Figure Lengend Snippet: A and B. Bel/Fu cells were pretreated NC, 5-Fu, Met and Com separately. After 48 h of treatment, flow cytometry was performed to analyze the rate of cell apoptosis. C. TUNEL assay was used to detect DNA damage after the different treatments. D. The protein levels of cl-caspase3, cl-PARP, Bax, and Bcl-2 were detected by western blotting. E and F. 24 h after different treatments, flow cytometry was performed to analyze the progression of the cell cycle. G. The protein levels of CDK4, cyclin D1, c-myc and p21 were detected by western blotting. **P < 0.01.

Article Snippet: Antibodies against caspase3, cleaved-caspase3 (cl-caspase3), PARP, cleaved-PARP (cl-PARP), Bax, Bcl-2, p-YAP, YAP, p-AMPK, AMPK, Akt, p-Akt, PTEN, HIF-1a, p21, c-myc and GAPDH were obtained from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA), the P-gp and MRP1 antibodies were obtained from Thermo Fisher Scientific Inc. (Rockford, IL, USA), and the cyclin D1 antibody was purchased from Abgent (San Diego, CA, USA).

Techniques: Flow Cytometry, TUNEL Assay, Western Blot

Journal: Cancer epidemiology, biomarkers & prevention : a publication of the American Association for Cancer Research, cosponsored by the American Society of Preventive Oncology

Article Title: The EARLY DETECTION RESEARCH NETWORK: A National Infrastructure to Support the Discovery, Development and Validation of Cancer Biomarkers

doi: 10.1158/1055-9965.EPI-20-0237

Figure Lengend Snippet: Tests in Clinical Laboratory Improvement Amendments (CLIA) Laboratories

Article Snippet: Veracyte Inc. Esoguard (methylated vimentin and cyclin A1) Detection of Barrett’s esophagus Sandford Markowitz, M.D.

Techniques: Biomarker Discovery, Expressing, Methylation, Diagnostic Assay, Multiplex Assay, Blocking Assay