Journal: BMC Cancer

Article Title: Non-endometrioid and high-grade endometrioid endometrial cancers show DNA fragmentation factor 40 (DFF40) and B-cell lymphoma 2 protein (BCL2) underexpression, which predicts disease-free and overall survival, but not DNA fragmentation factor 45 (DFF45) underexpression

doi: 10.1186/s12885-018-4333-6

Figure Lengend Snippet: DFF40*, DFF45**, and BCL2*** expression (× 200) in different histological types of endometrial cancers. *DNA fragmentation factor 40; **DNA fragmentation factor 45; ***B-cell lymphoma 2 protein; $ Endometrial cancer

Article Snippet: Immunohistochemical staining was performed with a rabbit polyclonal antibody against DFF40 (Abcam, Cambridge, UK) diluted 1:50, a monoclonal mouse anti-human antibody against BCL2 (Leica Biosystems Newcastle Ltd., Newcastle Upon Tyne, UK) diluted 1:200 and a rabbit polyclonal antibody against DFF45 (Abcam, Cambridge, UK) diluted 1:100 using a previously described protocol [ , ].

Techniques: Expressing

Journal: BMC Cancer

Article Title: Non-endometrioid and high-grade endometrioid endometrial cancers show DNA fragmentation factor 40 (DFF40) and B-cell lymphoma 2 protein (BCL2) underexpression, which predicts disease-free and overall survival, but not DNA fragmentation factor 45 (DFF45) underexpression

doi: 10.1186/s12885-018-4333-6

Figure Lengend Snippet: Immunoexpression of DFF40 * , DFF45 ** , and BCL2 *** in endometrial cancers in relation to histological type and study groups A and B

Article Snippet: Immunohistochemical staining was performed with a rabbit polyclonal antibody against DFF40 (Abcam, Cambridge, UK) diluted 1:50, a monoclonal mouse anti-human antibody against BCL2 (Leica Biosystems Newcastle Ltd., Newcastle Upon Tyne, UK) diluted 1:200 and a rabbit polyclonal antibody against DFF45 (Abcam, Cambridge, UK) diluted 1:100 using a previously described protocol [ , ].

Techniques:

Journal: BMC Cancer

Article Title: Non-endometrioid and high-grade endometrioid endometrial cancers show DNA fragmentation factor 40 (DFF40) and B-cell lymphoma 2 protein (BCL2) underexpression, which predicts disease-free and overall survival, but not DNA fragmentation factor 45 (DFF45) underexpression

doi: 10.1186/s12885-018-4333-6

Figure Lengend Snippet: Relative risks (RRs) and 95% confidence intervals (CIs) of recurrence (R+), lymph node involvement (LN+), lymphovascular space invasion (LVSI+), and myometrial invasion over 50% (MI > 50%) in patients with DFF40*-, DFF45**-, and BCL2***-negative endometrial cancers

Article Snippet: Immunohistochemical staining was performed with a rabbit polyclonal antibody against DFF40 (Abcam, Cambridge, UK) diluted 1:50, a monoclonal mouse anti-human antibody against BCL2 (Leica Biosystems Newcastle Ltd., Newcastle Upon Tyne, UK) diluted 1:200 and a rabbit polyclonal antibody against DFF45 (Abcam, Cambridge, UK) diluted 1:100 using a previously described protocol [ , ].

Techniques:

Journal: BMC Cancer

Article Title: Non-endometrioid and high-grade endometrioid endometrial cancers show DNA fragmentation factor 40 (DFF40) and B-cell lymphoma 2 protein (BCL2) underexpression, which predicts disease-free and overall survival, but not DNA fragmentation factor 45 (DFF45) underexpression

doi: 10.1186/s12885-018-4333-6

Figure Lengend Snippet: Kaplan-Meier survival curves for overall and disease-free survival in DFF40*-, DFF45**-, and BCL2***-negative and positive cases in the entire study cohort and in Groups A $ and B $$ , respectively. *DFF40 - DNA fragmentation factor 40; **DFF45 - DNA fragmentation factor 45; ***BCL2 - B-cell lymphoma 2; $ Group A – includes low- and moderate-grade endometroid endometrial cancers; $$ Group B – includes non-endometroid and high-grade endometroid endometrial cancers; # p -value statistically significant

Article Snippet: Immunohistochemical staining was performed with a rabbit polyclonal antibody against DFF40 (Abcam, Cambridge, UK) diluted 1:50, a monoclonal mouse anti-human antibody against BCL2 (Leica Biosystems Newcastle Ltd., Newcastle Upon Tyne, UK) diluted 1:200 and a rabbit polyclonal antibody against DFF45 (Abcam, Cambridge, UK) diluted 1:100 using a previously described protocol [ , ].

Techniques:

Journal: Archives of Medical Science : AMS

Article Title: Immunoexpression of DNA fragmentation factor 40, DNA fragmentation factor 45, and B-cell lymphoma 2 protein in normal human endometrium and uterine myometrium depends on menstrual cycle phase and menopausal status

doi: 10.5114/aoms.2017.69383

Figure Lengend Snippet: Expression of DFF40*, DFF45** and Bcl-2*** (200×) in normal human endometrium and myometrium *DNA fragmentation factor 40, **DNA fragmentation factor 45, ***B-cell lymphoma 2.

Article Snippet: A standard immunohistochemical technique was performed using a rabbit polyclonal antibody to DFF45 (Abcam, Cambridge, UK) at a dilution of 1 : 100, a rabbit polyclonal antibody to DFF40 (Abcam, Cambridge, UK) at a dilution of 1 : 50, and a monoclonal mouse anti-human antibody to Bcl-2 (Leica Microsystems GmbH, Wetzlar, Germany) at a dilution of 1 : 200.

Techniques: Expressing

Journal: Archives of Medical Science : AMS

Article Title: Immunoexpression of DNA fragmentation factor 40, DNA fragmentation factor 45, and B-cell lymphoma 2 protein in normal human endometrium and uterine myometrium depends on menstrual cycle phase and menopausal status

doi: 10.5114/aoms.2017.69383

Figure Lengend Snippet: .Median DFF40*, DFF45**, and Bcl-2*** expression in the normal endometrial glands, stroma, and uterine myometrium *DNA fragmentation factor 40, **DNA fragmentation factor 45, ***B-cell lymphoma 2, #p-value statistically significant. DFF40* expression

Article Snippet: A standard immunohistochemical technique was performed using a rabbit polyclonal antibody to DFF45 (Abcam, Cambridge, UK) at a dilution of 1 : 100, a rabbit polyclonal antibody to DFF40 (Abcam, Cambridge, UK) at a dilution of 1 : 50, and a monoclonal mouse anti-human antibody to Bcl-2 (Leica Microsystems GmbH, Wetzlar, Germany) at a dilution of 1 : 200.

Techniques: Expressing

Journal: Archives of Medical Science : AMS

Article Title: Immunoexpression of DNA fragmentation factor 40, DNA fragmentation factor 45, and B-cell lymphoma 2 protein in normal human endometrium and uterine myometrium depends on menstrual cycle phase and menopausal status

doi: 10.5114/aoms.2017.69383

Figure Lengend Snippet: Immunoexpression of DFF40; DFF45 and Bcl-2 in normal human endometrium and normal human uterine myometrium in different phases of menstrual cycle and postmenopausally

Article Snippet: A standard immunohistochemical technique was performed using a rabbit polyclonal antibody to DFF45 (Abcam, Cambridge, UK) at a dilution of 1 : 100, a rabbit polyclonal antibody to DFF40 (Abcam, Cambridge, UK) at a dilution of 1 : 50, and a monoclonal mouse anti-human antibody to Bcl-2 (Leica Microsystems GmbH, Wetzlar, Germany) at a dilution of 1 : 200.

Techniques:

Journal: Nucleic Acids Research

Article Title: Engineered apoptotic nucleases for chromatin research

doi: 10.1093/nar/gkm486

Figure Lengend Snippet: Inserting TEVP cleavage sites downstream of caspase-3 cleavage sites within DFF45 is effective in generating DFF40 nuclease activity to be dependent on TEVP cleavage. ( A and D ) Amino acid sequence of mouse DFF45 (fragments containing both protease cleavage sites). The caspase-3 recognition sequences are underlined, the TEVP recognition sequences are in italics. ( B and E ) The indicated forms of mouse DFF heterodimers, either wild-type (WT), swap (S) or insertion (I) mutants, were expressed in E. coli , purified and treated either with caspase-3, TEVP, both or neither. Western blot of a SDS–PAGE gel probed with anti-DFF45 antibodies (B) or with a mixture of anti-DFF40 and anti-DFF45 antibodies (E); arrowheads mark the positions of intact DFF45 and GST-DFF40. ( C and F ) Agarose gel of plasmid DNA substrate incubated with the indicated proteinase-treated DFF species, either wild-type (WT), swap (S) or insertion (I) mutants.

Article Snippet: Membrane-immoblilized proteins were probed with the following commercial antibodies: rabbit anti-human DFF40 polyclonal antibodies and/or rabbit anti-human DFF45 N-terminus polyclonal antibodies (Pharmingen).

Techniques: Activity Assay, Sequencing, Purification, Western Blot, SDS Page, Agarose Gel Electrophoresis, Plasmid Preparation, Incubation

Journal: Nucleic Acids Research

Article Title: Engineered apoptotic nucleases for chromatin research

doi: 10.1093/nar/gkm486

Figure Lengend Snippet: Expression of DFF-T in yeast. ( A ) Co-expression of TEVP, GST-DFF40 and the TEVP-cleavable DFF45 I 1 I 2 , is lethal in yeast cells. Suspended yeast cells with or without an integrated TEVP gene and harboring the indicated forms of mouse DFF in a yeast expression plasmid were serially diluted 10-fold, and 5 μl aliquots were spotted onto galactose plates. Plates were photographed after 3 days of growth at 30°C. ( B ) Nucleosomal DNA laddering in vivo in yeast requires the combined expression of TEVP, GST-DFF40 and the DFF45 I 1 I 2 mutant. Agarose gel of genomic DNA purified from yeast harboring the indicated forms of mouse DFF in a yeast expression plasmid after 6 h of galactose induction.

Article Snippet: Membrane-immoblilized proteins were probed with the following commercial antibodies: rabbit anti-human DFF40 polyclonal antibodies and/or rabbit anti-human DFF45 N-terminus polyclonal antibodies (Pharmingen).

Techniques: Expressing, Plasmid Preparation, DNA Laddering, In Vivo, Mutagenesis, Agarose Gel Electrophoresis, Purification

Journal: Nucleic Acids Research

Article Title: Engineered apoptotic nucleases for chromatin research

doi: 10.1093/nar/gkm486

Figure Lengend Snippet: High-level expression from synthetic human DFF genes in E. coli . ( A ) Coomassie blue-stained SDS–PAGE gel of total cell protein from uninduced or induced E. coli BL21 (DE3) transformants, both carrying DFF40 synthetic genes, and either DFF45 wild-type (DFF-wt) or the DFF45I 1 I 2 mutant (DFF-T) synthetic genes, in the pETDuet™ expression plasmid. ( B ) Agarose gel of plasmid DNA substrate incubated with the indicated proteinase-treated DFF nucleases.

Article Snippet: Membrane-immoblilized proteins were probed with the following commercial antibodies: rabbit anti-human DFF40 polyclonal antibodies and/or rabbit anti-human DFF45 N-terminus polyclonal antibodies (Pharmingen).

Techniques: Expressing, Staining, SDS Page, Mutagenesis, Plasmid Preparation, Agarose Gel Electrophoresis, Incubation