Journal: Frontiers in Genetics

Article Title: Post-transcriptional regulation of BRCA1 through its coding sequence by the miR-15/107 group of miRNAs

doi: 10.3389/fgene.2015.00242

Figure Lengend Snippet: Putative miR-15/107 targets within the BRCA1 CDS. (A) Schematic representation of protein-coding BRCA1 transcript. (B) Density function and cumulative distribution of CDS length across all human coding mRNAs. (C) The rna22-predicted target site for the miR-15/107 group together with the corresponding miRNA:mRNA heteroduplexes.

Article Snippet: Primary antibodies include anti-BRCA1 (D54A8, Cell Signaling, Danvers, MA, USA) and anti-β-actin (8H10D10, Cell Signaling).

Techniques:

Journal: Frontiers in Genetics

Article Title: Post-transcriptional regulation of BRCA1 through its coding sequence by the miR-15/107 group of miRNAs

doi: 10.3389/fgene.2015.00242

Figure Lengend Snippet: Suppression of BRCA1 mRNA by miR-15/107 miRNAs. Cell lines transfected with miR-15/107 miRNA precursors show a significant decrease in the amount of available BRCA1 transcript 48 h post-transfection as detected by qRT-PCR. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 compared to control scramble miR by Student’s t -test, n = 3 in each group.

Article Snippet: Primary antibodies include anti-BRCA1 (D54A8, Cell Signaling, Danvers, MA, USA) and anti-β-actin (8H10D10, Cell Signaling).

Techniques: Transfection, Quantitative RT-PCR, Control

Journal: Frontiers in Genetics

Article Title: Post-transcriptional regulation of BRCA1 through its coding sequence by the miR-15/107 group of miRNAs

doi: 10.3389/fgene.2015.00242

Figure Lengend Snippet: Characterization of a novel miR-15/107 target site within the BRCA1 CDS. The putative binding site for the miR-15/107 group in BRCA1’s CDS was cloned into a luciferase reporter vector along with a corresponding construct containing mutations within the putative miRNA ‘seed’ recognition sites to confirm specificity. ∗ P < 0.05, ∗∗ P < 0.01 compared to control scramble target sequence by Student’s t -test, n = 3 in each group.

Article Snippet: Primary antibodies include anti-BRCA1 (D54A8, Cell Signaling, Danvers, MA, USA) and anti-β-actin (8H10D10, Cell Signaling).

Techniques: Binding Assay, Clone Assay, Luciferase, Plasmid Preparation, Construct, Control, Sequencing

Journal: Frontiers in Genetics

Article Title: Post-transcriptional regulation of BRCA1 through its coding sequence by the miR-15/107 group of miRNAs

doi: 10.3389/fgene.2015.00242

Figure Lengend Snippet: Sequestration of miR-15/107 miRNAs impacts BRCA1 mRNA. Cells transfected with luciferase reporter plasmids containing antisense (as) miRNA sequences or predicted wild type (WT) BRCA1 miR-15/107 MRE demonstrate varying levels of BRCA1 expression rescue. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 compared to control by Student’s t -test, n = 3 in each group.

Article Snippet: Primary antibodies include anti-BRCA1 (D54A8, Cell Signaling, Danvers, MA, USA) and anti-β-actin (8H10D10, Cell Signaling).

Techniques: Transfection, Luciferase, Expressing, Control

Journal: Frontiers in Genetics

Article Title: Post-transcriptional regulation of BRCA1 through its coding sequence by the miR-15/107 group of miRNAs

doi: 10.3389/fgene.2015.00242

Figure Lengend Snippet: Translational suppression of BRCA1 by miR-15/107 miRNAs. Western immunoblots from hTERT-HPNE, HCT-116, and MIA PaCa-2 cells demonstrate relative levels of BRCA1 protein expression at 72 h post-transfection with control scramble miRNA and anti-miR precursors, miRNA and anti-miR precursors, or siBRCA1 control.

Article Snippet: Primary antibodies include anti-BRCA1 (D54A8, Cell Signaling, Danvers, MA, USA) and anti-β-actin (8H10D10, Cell Signaling).

Techniques: Western Blot, Expressing, Transfection, Control

Journal: eLife

Article Title: LIN37-DREAM prevents DNA end resection and homologous recombination at DNA double-strand breaks in quiescent cells

doi: 10.7554/eLife.68466

Figure Lengend Snippet:

Article Snippet: Antibody , Anti-BRCA1 (Mouse monoclonal) , R and D Systems , Custom made (Andre Nussenzweig, NCI) , WB (1:1000); for mouse BRCA1.

Techniques: Recombinant, Plasmid Preparation, Genome Wide, CRISPR, Clone Assay, Flow Cytometry, Staining, Sequencing, Software, Microscopy

Journal: Scientific Reports

Article Title: TP53 supports basal-like differentiation of mammary epithelial cells by preventing translocation of deltaNp63 into nucleoli

doi: 10.1038/srep04663

Figure Lengend Snippet: (a) Immunofluorescence showing the ΔNp63 protein (top row) in MCF7 cells treated as indicated above. BRCA1 protein (second row) is readily detectable in control cells, but disappears after treatments promoting expression of ΔNp63. MDM2 protein (third row) correlates with the expression of BRCA1. (b) qRT-PCR showing fold change in mRNA levels for ΔNp63, MDM2, and BRCA1 following treatments indicated above. (c) Western blot confirming an effective reduction of BRCA1 protein level and stabilization of the TP53 protein after treatments indicated above. F12, cells were cultured in DMEM/F12 medium instead of the regular DMEM; Serdemetan, cells treated with MDM2 inhibitor as described in Materials and Methods; siBRCA1 and siMDM2, cells treated with siRNAs targeting corresponding genes. Scale bars correspond to 20 µm.

Article Snippet: Primary antibodies were used as follows: goat anti-TAp63 (Santa Cruz, sc-8608, diluted 1:50), rabbit anti-Ki67 (Abcam, ab15580, diluted 1:200), goat anti-ΔNp63 (Santa Cruz, sc-8609, diluted 1:100), rabbit anti-MDM2 (Abcam, ab58530, diluted 1:100), mouse monoclonal anti-Krt18 (Abcam, ab668, diluted 1:100), mouse monoclonal anti-p53 (Abcam, ab26, diluted 1:100), mouse monoclonal anti-p53 (Abcam, PAb 240, diluted 1:100), mouse anti-human BRCA1 (Calbiochem, OP92, diluted 1:100), and mouse monoclonal anti-Nucleophosmin (Invitrogen, FC-61991, diluted 1:200, a kind gift of Karita Peltonen and Prof. Marikki Laiho).

Techniques: Immunofluorescence, Control, Expressing, Quantitative RT-PCR, Western Blot, Cell Culture

Journal: Scientific Reports

Article Title: TP53 supports basal-like differentiation of mammary epithelial cells by preventing translocation of deltaNp63 into nucleoli

doi: 10.1038/srep04663

Figure Lengend Snippet: (a) Untreated MCF10A cells express high levels of ΔNp63, and low levels of BRCA1 and MDM2. Cells plated at a low density translocate ΔNp63 to nucleoli, and elevate expression of BRCA1, and MDM2. (b) Knockdown of TP63 (siTP63) results in upregulation of BRCA1 and MDM2. (c) Western blot confirming an efficient siRNA-mediated knockdown of BRCA1 and ΔNp63 proteins, and lack of TP53 in TP53 −/− MCF10A cells. Notice a higher level of ΔNp63 protein in BRCA1-depleted TP53-mutant cells. Asterisk indicates a 75 kDa band from a molecular weight marker loaded in the same lane as the siMDM2 sample. (d) TP53-deficiency in MCF10A cells leads to an increase in MDM2 mRNA level as measured by qRT-PCR. ( e ) Immunofluorescence staining of TP53 −/− and its parental isogenic TP53 +/+ MCF10A cell lines demonstrates a ubiquitous relocation of ΔNp63 protein from the nucleoplasm in TP53 +/+ cells (arrowheads) to nucleoli in TP53 −/− cells (arrows), and upregulation of BRCA1 in TP53 −/− cells. Insets illustrate the differences at a higher magnification. Scale bars correspond to 20 µm.

Article Snippet: Primary antibodies were used as follows: goat anti-TAp63 (Santa Cruz, sc-8608, diluted 1:50), rabbit anti-Ki67 (Abcam, ab15580, diluted 1:200), goat anti-ΔNp63 (Santa Cruz, sc-8609, diluted 1:100), rabbit anti-MDM2 (Abcam, ab58530, diluted 1:100), mouse monoclonal anti-Krt18 (Abcam, ab668, diluted 1:100), mouse monoclonal anti-p53 (Abcam, ab26, diluted 1:100), mouse monoclonal anti-p53 (Abcam, PAb 240, diluted 1:100), mouse anti-human BRCA1 (Calbiochem, OP92, diluted 1:100), and mouse monoclonal anti-Nucleophosmin (Invitrogen, FC-61991, diluted 1:200, a kind gift of Karita Peltonen and Prof. Marikki Laiho).

Techniques: Expressing, Knockdown, Western Blot, Mutagenesis, Molecular Weight, Marker, Quantitative RT-PCR, Immunofluorescence, Staining

Journal: Scientific Reports

Article Title: TP53 supports basal-like differentiation of mammary epithelial cells by preventing translocation of deltaNp63 into nucleoli

doi: 10.1038/srep04663

Figure Lengend Snippet: TP53 prevents translocation of ΔNp63 from the nucleoplasm into nucleoli. Active TP53 and a nucleoplasmic ΔNp63 inversely correlate with MDM2, BRCA1, and TAp63. MDM2 and BRCA1 in luminal cells inhibit TP53, thus preventing the nucleoplasmic expression of ΔNp63 and suppressing the basal-like differentiation. Loss of TP53 leads to an upregulation of TAp63, which is associated with an unlimited cell proliferation rather than a particular epithelial subtype. Relocation of ΔNp63 to nucleoli in basal epithelial cells is associated with EMT.

Article Snippet: Primary antibodies were used as follows: goat anti-TAp63 (Santa Cruz, sc-8608, diluted 1:50), rabbit anti-Ki67 (Abcam, ab15580, diluted 1:200), goat anti-ΔNp63 (Santa Cruz, sc-8609, diluted 1:100), rabbit anti-MDM2 (Abcam, ab58530, diluted 1:100), mouse monoclonal anti-Krt18 (Abcam, ab668, diluted 1:100), mouse monoclonal anti-p53 (Abcam, ab26, diluted 1:100), mouse monoclonal anti-p53 (Abcam, PAb 240, diluted 1:100), mouse anti-human BRCA1 (Calbiochem, OP92, diluted 1:100), and mouse monoclonal anti-Nucleophosmin (Invitrogen, FC-61991, diluted 1:200, a kind gift of Karita Peltonen and Prof. Marikki Laiho).

Techniques: Translocation Assay, Expressing

Journal: Molecular cell

Article Title: Restoration of replication fork stability in BRCA1- and BRCA2-deficient cells by inactivation of SNF2-family fork remodelers

doi: 10.1016/j.molcel.2017.09.036

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit polyclonal anti-BRCA1 , Bethyl Laboratories , Cat#A301-378A; RRID:AB_937737.

Techniques: Virus, Subcloning, Recombinant, Transfection, Protease Inhibitor, In Situ, Microscopy, Plasmid Preparation, Sequencing, shRNA, Software, Irradiation