breast cancer tissue microarray bc081115  (Biomax Inc)

Journal: The EMBO Journal

Article Title: A novel non-coding RNA lncRNA-JADE connects DNA damage signalling to histone H4 acetylation

doi: 10.1038/emboj.2013.221

Figure Lengend Snippet: ATM-dependent regulation of lncRNA expression in response to DNA damage. (A) Experimental layout to identify ATM-dependent lncRNAs. Atm+/+ and Atm−/− mouse embryonic fibroblasts (MEFs) were treated with NCS (200 ng/ml) and harvested at indicated time points for microarray analyses. (B) The number of ATM-dependent lncRNAs upon DNA damage. (C) A representative group of ATM-dependent and DNA damage-induced lncRNAs. Green or red colour on the heat map indicates a decrease or an increase in the lncRNA level and colour intensities correspond to relative signal levels on a logarithmic scale. (D) Quantitative PCR validation of representative lncRNAs. Data represent the mean of three experimental replicates, with error bars depicting s.d.

Article Snippet: LncRNA in situ hybridization Breast cancer tissue microarray (no. {"type":"entrez-nucleotide","attrs":{"text":"BC081115","term_id":"51703523","term_text":"BC081115"}} BC081115 ) was purchased from Biomax, including 100 breast tumour samples and 10 control breast tissue samples.

Techniques: Expressing, Microarray, Real-time Polymerase Chain Reaction, Biomarker Discovery

Journal: The EMBO Journal

Article Title: A novel non-coding RNA lncRNA-JADE connects DNA damage signalling to histone H4 acetylation

doi: 10.1038/emboj.2013.221

Figure Lengend Snippet: LncRNA-JADE is induced after DNA damage. (A) Schematic illustration showing Jade1 and lncRNA-JADE genes in mouse and human. (B) Mouse and human lncRNA-JADE are induced in an ATM-dependent manner after DNA damage. (C) DNA damage positively regulates lncRNA-JADE promoter activity in an ATM-dependent manner. ATM-IN: ATM inhibitor. (D) Schematic illustration showing the NF-κB binding elements in the Jade1 promoter. DNA damage induces the activity of lncRNA-JADE promoter in an NF-κB-dependent manner. NF-κB-IN: NF-κB inhibitor. (E) Expression of LncRNA-JADE is regulated by ATM and NF-κB after DNA damage. IRF-1 (interferon response factor-1) is served as a positive control in the NF-κB signalling. Graphic data in this figure present the mean of three experimental replicates and error bars depict s.d.

Article Snippet: LncRNA in situ hybridization Breast cancer tissue microarray (no. {"type":"entrez-nucleotide","attrs":{"text":"BC081115","term_id":"51703523","term_text":"BC081115"}} BC081115 ) was purchased from Biomax, including 100 breast tumour samples and 10 control breast tissue samples.

Techniques: Activity Assay, Binding Assay, Expressing, Positive Control

Journal: The EMBO Journal

Article Title: A novel non-coding RNA lncRNA-JADE connects DNA damage signalling to histone H4 acetylation

doi: 10.1038/emboj.2013.221

Figure Lengend Snippet: LncRNA-JADE positively regulates histone H4 acetylation through Jade1. (A) H4 acetylation is induced after DNA damage. MCF7 cells were treated with NCS (500 ng/ml). H4Ac: total histone H4 acetylation; K5, K8, K12: Histone H4 acetylation at lysine 5, 8, or 12. (B) Jade1 knockdown abolishes the induction of H4 acetylation after DNA damage. (C) LncRNA-JADE positively regulates H4 acetylation and Jade1. Overexpression of lncRNA-JADE enhanced the induction of H4 acetylation and Jade1, and knockdown of lncRNA-JADE abolished the induction of H4 acetylation and Jade1. Semi-quantification of proteins is shown at the bottom.

Article Snippet: LncRNA in situ hybridization Breast cancer tissue microarray (no. {"type":"entrez-nucleotide","attrs":{"text":"BC081115","term_id":"51703523","term_text":"BC081115"}} BC081115 ) was purchased from Biomax, including 100 breast tumour samples and 10 control breast tissue samples.

Techniques: Knockdown, Over Expression

Journal: The EMBO Journal

Article Title: A novel non-coding RNA lncRNA-JADE connects DNA damage signalling to histone H4 acetylation

doi: 10.1038/emboj.2013.221

Figure Lengend Snippet: Brca1 binds lncRNA-JADE and mediates Jade1 induction in the DNA damage response via p300-containing transcription complex. (A) Schematic illustration showing the p300/CBP binding elements in the Jade1 promoter. (B) p300 physically interacts with the promoter region of Jade1 gene. Control or lncRNA-JADE knockdown MCF7 cells were treated with or without NCS (200 ng/ml) and cell lysates were immunoprecipitated with control IgG or p300 antibodies. The p300-binding activity of Jade1 promoter DNA was quantified by qPCR. (C) Brca1 interacts with p300 and this interaction is increased after DNA damage. (D) Jade1 promoter activity is induced in a Brca1-dependent manner after DNA damage. MCF-7 cells were infected with lentiviruses expressing control or Brca1 shRNA. The cells were transfected with pGL3-control vector (SV40 promoter) or Jade1 promoter-driven firefly luciferase expression vector and Renilla luciferase expression vector 2 days post infection. They were treated with NCS (500 ng/ml) 24 h after transfection and then harvested 16 h after treatment. Firefly luciferase activity was measured and normalized to the activity of Renilla luciferase. (E) LncRNA-JADE physically interacts with Brca1. 5′- and 3′-deletion mutants of lncRNA-JADE were generated as indicated. Two pairs of primers were used to detect the Brca1-binding sequences of lncRNA-JADE in RIP assays. Graphic data in this figure present the mean of three experimental replicates and error bars depict s.d.

Article Snippet: LncRNA in situ hybridization Breast cancer tissue microarray (no. {"type":"entrez-nucleotide","attrs":{"text":"BC081115","term_id":"51703523","term_text":"BC081115"}} BC081115 ) was purchased from Biomax, including 100 breast tumour samples and 10 control breast tissue samples.

Techniques: Binding Assay, Control, Knockdown, Immunoprecipitation, Activity Assay, Infection, Expressing, shRNA, Transfection, Plasmid Preparation, Luciferase, Generated

Journal: The EMBO Journal

Article Title: A novel non-coding RNA lncRNA-JADE connects DNA damage signalling to histone H4 acetylation

doi: 10.1038/emboj.2013.221

Figure Lengend Snippet: Biological functions of lncRNA-JADE in human MCF7 cells. (A) LncRNA-JADE positively regulates MCF7 cell proliferation. (B) Altering lncRNA-JADE expression affects DNA damage-induced cell-cycle arrest. Cell-cycle profiles were analysed by flow cytometry using propidium iodide-stained cells. (C) Knockdown of lncRNA-JADE increases cell apoptosis in the control and NCS-treated cells. The percentage of TUNEL-positive cells was summarized in the graph. (D) Knockdown of lncRNA-JADE increases the cell sensitivity to DNA damaging drugs NCS, Etopside, and Bleomycin. MCF7 cells were treated with DNA damaging agents as indicated and cultured for 48 h and cell viability was measured. Graphic data in this figure present the mean of three biological replicates and error bars depict s.d.

Article Snippet: LncRNA in situ hybridization Breast cancer tissue microarray (no. {"type":"entrez-nucleotide","attrs":{"text":"BC081115","term_id":"51703523","term_text":"BC081115"}} BC081115 ) was purchased from Biomax, including 100 breast tumour samples and 10 control breast tissue samples.

Techniques: Expressing, Flow Cytometry, Staining, Knockdown, Control, TUNEL Assay, Cell Culture

Journal: The EMBO Journal

Article Title: A novel non-coding RNA lncRNA-JADE connects DNA damage signalling to histone H4 acetylation

doi: 10.1038/emboj.2013.221

Figure Lengend Snippet: Knockdown of lncRNA-JADE inhibits mammary tumour growth in vivo. (A) Higher levels of lncRNA-JADE in human breast cancer tissues in comparison with normal breast tissues. In situ hybridization of lncRNA-JADE was performed on tissue microarray comprised of human normal breast and breast cancer tissues. (B) Correlation between lncRNA-JADE and Jade1 up-expression in breast cancer tissue cDNA array. The level of lncRNA-JADE and Jade1 was measured by RT–PCR. The lncRNA-JADE expression demonstrated a significant correlation with the Jade1 expression according to the Spearman correlation coefficient (r=0.6983 and P=0.0294). (C) Comparison of survival curves between patients with Jade1 overexpression and patients with normal Jade1 expression using TCGA data in breast invasive carcinomas. (D) Knockdown of lncRNA-JADE inhibits xenografted 4T1 tumour growth in vivo. One million luciferase expressing 4T1 cells stably expressing control or lncRNA-JADE shRNA were injected into the mammary fat pad of each Balb/cSCID mouse. Two weeks after injection, luciferase activity was measured and quantified by an IVIS device (left panel and upper right panel). Breast tumour size was measured in the mice for 24 days (bottom right panel). Graphic data present the mean of five mice and error bars depict s.d.

Article Snippet: LncRNA in situ hybridization Breast cancer tissue microarray (no. {"type":"entrez-nucleotide","attrs":{"text":"BC081115","term_id":"51703523","term_text":"BC081115"}} BC081115 ) was purchased from Biomax, including 100 breast tumour samples and 10 control breast tissue samples.

Techniques: Knockdown, In Vivo, Comparison, In Situ Hybridization, Microarray, Expressing, Reverse Transcription Polymerase Chain Reaction, Over Expression, Luciferase, Stable Transfection, Control, shRNA, Injection, Activity Assay

Journal: The EMBO Journal

Article Title: A novel non-coding RNA lncRNA-JADE connects DNA damage signalling to histone H4 acetylation

doi: 10.1038/emboj.2013.221

Figure Lengend Snippet: ATM-dependent regulation of lncRNA expression in response to DNA damage. (A) Experimental layout to identify ATM-dependent lncRNAs. Atm+/+ and Atm−/− mouse embryonic fibroblasts (MEFs) were treated with NCS (200 ng/ml) and harvested at indicated time points for microarray analyses. (B) The number of ATM-dependent lncRNAs upon DNA damage. (C) A representative group of ATM-dependent and DNA damage-induced lncRNAs. Green or red colour on the heat map indicates a decrease or an increase in the lncRNA level and colour intensities correspond to relative signal levels on a logarithmic scale. (D) Quantitative PCR validation of representative lncRNAs. Data represent the mean of three experimental replicates, with error bars depicting s.d.

Article Snippet: Breast cancer tissue microarray (no. {"type":"entrez-nucleotide","attrs":{"text":"BC081115","term_id":"51703523","term_text":"BC081115"}} BC081115 ) was purchased from Biomax, including 100 breast tumour samples and 10 control breast tissue samples.

Techniques: Expressing, Microarray, Real-time Polymerase Chain Reaction, Biomarker Discovery

Journal: The EMBO Journal

Article Title: A novel non-coding RNA lncRNA-JADE connects DNA damage signalling to histone H4 acetylation

doi: 10.1038/emboj.2013.221

Figure Lengend Snippet: Knockdown of lncRNA-JADE inhibits mammary tumour growth in vivo. (A) Higher levels of lncRNA-JADE in human breast cancer tissues in comparison with normal breast tissues. In situ hybridization of lncRNA-JADE was performed on tissue microarray comprised of human normal breast and breast cancer tissues. (B) Correlation between lncRNA-JADE and Jade1 up-expression in breast cancer tissue cDNA array. The level of lncRNA-JADE and Jade1 was measured by RT–PCR. The lncRNA-JADE expression demonstrated a significant correlation with the Jade1 expression according to the Spearman correlation coefficient (r=0.6983 and P=0.0294). (C) Comparison of survival curves between patients with Jade1 overexpression and patients with normal Jade1 expression using TCGA data in breast invasive carcinomas. (D) Knockdown of lncRNA-JADE inhibits xenografted 4T1 tumour growth in vivo. One million luciferase expressing 4T1 cells stably expressing control or lncRNA-JADE shRNA were injected into the mammary fat pad of each Balb/cSCID mouse. Two weeks after injection, luciferase activity was measured and quantified by an IVIS device (left panel and upper right panel). Breast tumour size was measured in the mice for 24 days (bottom right panel). Graphic data present the mean of five mice and error bars depict s.d.

Article Snippet: Breast cancer tissue microarray (no. {"type":"entrez-nucleotide","attrs":{"text":"BC081115","term_id":"51703523","term_text":"BC081115"}} BC081115 ) was purchased from Biomax, including 100 breast tumour samples and 10 control breast tissue samples.

Techniques: Knockdown, In Vivo, Comparison, In Situ Hybridization, Microarray, Expressing, Reverse Transcription Polymerase Chain Reaction, Over Expression, Luciferase, Stable Transfection, Control, shRNA, Injection, Activity Assay