Journal: Oncotarget

Article Title: Poly (ADP-ribose) polymerase 1 transcriptional regulation: A novel crosstalk between histone modification H3K9ac and ETS1 motif hypomethylation in BRCA1-mutated ovarian cancer

doi:

Figure Lengend Snippet: (A) the schematic represents the cytosine located at the ETS1 and ETS2 motif that were point mutated to generate the thymine. (Bi-Biv) 293T cells, SKOV3 cells and primary non-mutated and BRCA1-mutated ovarian cancer cells were transfected with mutant plasmids. At 24 hours after transfection, whole-cell extracts were analyzed for luciferase activity. Bar graphs show mean ± SD. *P < 0.05 vs. Control. Mut., Mutation. (Ci) the schematic represents the selected nucleotide sequence with or without a methyl group at the fifth position of the cytosine pyrimidine ring at the ETS1 motif. (Cii) the CD spectra of the selected nucleotide sequence in the presence of 100 mM Na+ or 100 mM K+ are shown.

Article Snippet: The SHRNA lentiviral particles of GCN5 (sc-37946-V), PCAF (sc-36198-V) and ETS1 (sc-29309-V) were purchased from Santa Cruz Biotechnology (CA, USA).

Techniques: Transfection, Mutagenesis, Luciferase, Activity Assay, Control, Sequencing, Circular Dichroism

Journal: Oncotarget

Article Title: Poly (ADP-ribose) polymerase 1 transcriptional regulation: A novel crosstalk between histone modification H3K9ac and ETS1 motif hypomethylation in BRCA1-mutated ovarian cancer

doi:

Figure Lengend Snippet: (Ai) chromatin immunoprecipitation was performed using antibodies to H3K9ac, H3K18ac, H3K27ac, H3K4me1, H3K4me2, H3K4me3, H3K36me3, H3K79me, H3K9me, H3K9me2, H3K9me3, H3K27me, H3K27me2, and H3K27me3. PCR was performed for regions around the ETS1 motif. A negative control without antibodies was included for comparison. (Aii) representative results of three independent experiments are shown. (B) expression levels of the GCN5, PCAF and ETS1 factors in BRCA1-mutated ovarian cancer. Bar graphs show mean ± SD, *P < 0.05 vs. Control.

Article Snippet: The SHRNA lentiviral particles of GCN5 (sc-37946-V), PCAF (sc-36198-V) and ETS1 (sc-29309-V) were purchased from Santa Cruz Biotechnology (CA, USA).

Techniques: Chromatin Immunoprecipitation, Negative Control, Comparison, Expressing, Control

Journal: Oncotarget

Article Title: Poly (ADP-ribose) polymerase 1 transcriptional regulation: A novel crosstalk between histone modification H3K9ac and ETS1 motif hypomethylation in BRCA1-mutated ovarian cancer

doi:

Figure Lengend Snippet: (Ai) RT-PCR showing GCN5, PCAF and ETS1 factors levels before and after knockdown by SHRNA, and normalized to β-actin expression. (Aii) representative results of three independent experiments are shown. (Bi) EdU labeling showing proliferation of GCN5, PCAF and ETS1-silenced and control cells. Blue, Hoechst 33342 labeling of cell nuclei; Red, EdU labeling of nuclei of proliferative cells. (Bii) the EdU incorporation rate was expressed as the ratio of EdU positive cells to total Hoechst33342 positive cells. (Ci and Cii) analysis of histone modification H3K9ac and transcription factor ETS1 enrichment around the ETS1 motif after the deletion of GCN5, PCAF or ETS1 factors. (D) The interactions of ETS1 and GCN5 or PCAF were examined by the immunoprecipitation of cell extracts with an antibody to ETS1, and the co-immunoprecipitation of ETS1, GCN5 and PCAF by western blot analysis. Results of Fig.3 A-D were obtained in BRCA1-mutated ovarian cancer cells, and the same results were also obtained in 293T cells, SKOV3 cells, and non-BRCA1 mutated ovarian cancer cells (data not shown). (Ei-Ev) the PARP1 expression levels after deletion of H3K9ac and ETS1 enrichment around the ETS1 motif in 293T cells, SKOV3 cells, and primary non-mutated and BRCA1-mutated ovarian cancer cells. Bar graphs show mean ± SD. *P < 0.05 vs. Control.

Article Snippet: The SHRNA lentiviral particles of GCN5 (sc-37946-V), PCAF (sc-36198-V) and ETS1 (sc-29309-V) were purchased from Santa Cruz Biotechnology (CA, USA).

Techniques: Reverse Transcription Polymerase Chain Reaction, Knockdown, shRNA, Expressing, Labeling, Control, Modification, Immunoprecipitation, Western Blot

Journal: Oncotarget

Article Title: Poly (ADP-ribose) polymerase 1 transcriptional regulation: A novel crosstalk between histone modification H3K9ac and ETS1 motif hypomethylation in BRCA1-mutated ovarian cancer

doi:

Figure Lengend Snippet: The following variables were analyzed: age at diagnosis, menstruation, grade, pT, pN, pM, FIGO, CA125, ascites, residual tumor, and p53 and ETS1 methylation. Pre, premenopausal; Post, postmenopausal; FIGO, International Federation of Gynecology and Obstetrics; Pos, positive; Neg, negative; M, methylated; UM, unmethylated.

Article Snippet: The SHRNA lentiviral particles of GCN5 (sc-37946-V), PCAF (sc-36198-V) and ETS1 (sc-29309-V) were purchased from Santa Cruz Biotechnology (CA, USA).

Techniques: Biomarker Discovery, Methylation

Journal: PLoS ONE

Article Title: FBI-1 Enhances ETS-1 Signaling Activity and Promotes Proliferation of Human Colorectal Carcinoma Cells

doi: 10.1371/journal.pone.0098041

Figure Lengend Snippet: (A-D) Lovo cells were transfected with ETS-RE-Luc reporter. (A, C-D) Cells were stably transfected with FBI-1 expression vector or empty vector. (B) Cells were transfected with FBI-1 siRNA or control siRNA. (D) Cells were transfected with ETS-1 siRNA (D). Lovo cells were treated with 5 ng/ml HGF (A-D, F), or ARQ-197 (C). (E) Overexpression or knockdown of FBI-1 were examined by western blot. The luciferase values are the mean ± SE of three independent experiments with similar results. *P<0.05 versus with the empty vector or the FBI-1 vector (A-D); versus with the control siRNA or the FBI-1 siRNA (A-D); versus with or without HGF/ARQ-197.

Article Snippet: The siRNA vector targeted to ETS-1 was purchased from Santa Cruz Biotechnology.

Techniques: Transfection, Stable Transfection, Expressing, Plasmid Preparation, Control, Over Expression, Knockdown, Western Blot, Luciferase

Journal: PLoS ONE

Article Title: FBI-1 Enhances ETS-1 Signaling Activity and Promotes Proliferation of Human Colorectal Carcinoma Cells

doi: 10.1371/journal.pone.0098041

Figure Lengend Snippet: (A-B) Western blotting with various antibodies showed the overexpression of FBI-1 or specific knockdown effect of FBI-1 siRNA on the endogenous FBI-1 protein level. Lovo cells were harvested for WB assays and detected by anti-FBI-1 antibody, anti-MMP1 antibody, anti-MMP9 antibody, anti-u-PA antibody, anti-c-Met antibody, anti-ETS-1 antibody, or anti-GAPDH antibody.

Article Snippet: The siRNA vector targeted to ETS-1 was purchased from Santa Cruz Biotechnology.

Techniques: Western Blot, Over Expression, Knockdown

Journal: PLoS ONE

Article Title: FBI-1 Enhances ETS-1 Signaling Activity and Promotes Proliferation of Human Colorectal Carcinoma Cells

doi: 10.1371/journal.pone.0098041

Figure Lengend Snippet: (A) Lovo cells stably transfected with FBI-1 or empty vector were prepared and subjected to ChIP by using IgG antibody (negative control) or antibodies for ETS-1, FBI-1 and p53. The Immunoprecipitated DNA fragment was quantified by real-time PCR assay. (B) Lovo cells, which were stably transfected with FBI-1 siRNA, or control siRNA, were harvested for the ChIP assays. The ChIP assays were performed with IgG antibody (negative control) or antibodies for ETS-1, FBI-1 and p53. *P<0.05 versus the empty vector or the FBI-1 vector (A); or versus the control siRNA or the FBI-1 siRNA (B). The cloned promoter region of MMP1 is showed above the figure.

Article Snippet: The siRNA vector targeted to ETS-1 was purchased from Santa Cruz Biotechnology.

Techniques: Stable Transfection, Transfection, Plasmid Preparation, Negative Control, Immunoprecipitation, Real-time Polymerase Chain Reaction, Control, Clone Assay

Journal: PLoS ONE

Article Title: FBI-1 Enhances ETS-1 Signaling Activity and Promotes Proliferation of Human Colorectal Carcinoma Cells

doi: 10.1371/journal.pone.0098041

Figure Lengend Snippet: (A–D) Lovo cells were transfected with FBI-1 expression vector or empty vector. (A–B) Lovo cells were co-transfected with MMP1-Luc vector, p53 siRNA, or control siRNA, p53 vector or empty vector as indicated. Following transfection, cells were harvested for the luciferase assay. The values are the mean ± SE of three independent experiments with similar results. (C–D) Lovo cells were co-transfected with p53 vector or empty vector, p53 siRNA or control siRNA. The ChIP assays were performed with IgG of anti-ETS-1 antibody. ( E–F ) The luciferase activities changing of EBS-luc, p21-luc and ARE-luc upon FBI-1 and p53 overexpression and knocking down. The values are the mean ± SE of three independent experiments with similar results. *P<0.05 versus the empty vector or the FBI-1 vector (A–D), *P<0.05 versus the empty vector or the p53 vector (A, C), *P<0.05 versus the p53 siRNA vector or the control siRNA vector (B, D).

Article Snippet: The siRNA vector targeted to ETS-1 was purchased from Santa Cruz Biotechnology.

Techniques: Transfection, Expressing, Plasmid Preparation, Control, Luciferase, Over Expression

Journal: PLoS ONE

Article Title: FBI-1 Enhances ETS-1 Signaling Activity and Promotes Proliferation of Human Colorectal Carcinoma Cells

doi: 10.1371/journal.pone.0098041

Figure Lengend Snippet: (A–B) LoVo, (C–D) HR8348, and (E–F) HT29 cells were stably transfected with the plasmids. Then, relative cell numbers were determined by the MTT assay. Relative cell numbers (A and B) shown are Mean± SD of triplicate measurements and have been repeated 3 times with similar O.D. value results. *P<0.05 versus the empty vector or the FBI-1 vector (A, C, E), *P<0.05 versus the FBI-1 siRNA vector or the control siRNA vector (B, D, F).

Article Snippet: The siRNA vector targeted to ETS-1 was purchased from Santa Cruz Biotechnology.

Techniques: Stable Transfection, Transfection, MTT Assay, Plasmid Preparation, Control

Journal: PLoS ONE

Article Title: FBI-1 Enhances ETS-1 Signaling Activity and Promotes Proliferation of Human Colorectal Carcinoma Cells

doi: 10.1371/journal.pone.0098041

Figure Lengend Snippet: (A–B) Lovo cells were stably transfected with the FBI-1 expression vector or the empty vector, or the FBI-1 siRNA vector or the control siRNA vector. Colony number was shown in the photographs (B).*P<0.05 versus the empty vector or the FBI-1 vector (A–B), *P<0.05 versus the FBI-1 siRNA vector or the control siRNA vector (A–B).

Article Snippet: The siRNA vector targeted to ETS-1 was purchased from Santa Cruz Biotechnology.

Techniques: Stable Transfection, Transfection, Expressing, Plasmid Preparation, Control