Journal: Cancer Biology & Therapy

Article Title: Engrailed 1 overexpression as a potential prognostic marker in quintuple-negative breast cancer

doi: 10.1080/15384047.2018.1423913

Figure Lengend Snippet: EN1 mRNA expression is highly expressed in the TNBC subtype. (A) The top-ranked 20 genes differentially expressed in TNBC were identified by analyzing public microarray datasets utilized in our previous report. (B) EN1 mRNA expression in breast cancer was evaluated in a dataset of 939 samples from The Cancer Genome Atlas (TCGA). Bar indicates the median value. (C) EN1 upregulation was evaluated in a dataset of 825 samples from TCGA, and the presented figure was generated using cBioPortal software (www. cbioportal.org). (D) EN1 mRNA expression was quantified via quantitative real-time (qRT)-PCR in breast-cancer cell lines and expressed relative to the calculated EN1/HPRT expression ratio. N, normal epithelial; L, luminal A/B; H, HER2; TN, TNBC.

Article Snippet: The cells were then blocked with 3% non-fat milk in 0.1% TX-100 in PBS, incubated (overnight, 4°C) with an anti-EN1 primary antibody (#27, AbClon), and washed (30 min) with 0.1% TX-100 in PBS.

Techniques: Expressing, Microarray, Generated, Software, Quantitative RT-PCR

Journal: Cancer Biology & Therapy

Article Title: Engrailed 1 overexpression as a potential prognostic marker in quintuple-negative breast cancer

doi: 10.1080/15384047.2018.1423913

Figure Lengend Snippet: EN1 protein is localized to both the nucleus and cytoplasm in tissues from patients with breast cancer. (A) siRNA-mediated EN1 knockdown was evaluated by quantitative real-time (qRT)-PCR in parental or ectopic EN1-overexpressing HCC38 cells. NT, non-targeting. (B) To verify the specificity of synthesized (clone 27) EN1 antibody, EN1 levels were confirmed via immunoblot analysis of parental or ectopic EN1-overexpressing HCC38 cells. (C) Positive (left) and negative (right) EN1 protein expression was identified by immunohistochemical staining of breast-cancer tissue microarray blocks with the synthesized (clone 27) EN1 antibody.

Article Snippet: The cells were then blocked with 3% non-fat milk in 0.1% TX-100 in PBS, incubated (overnight, 4°C) with an anti-EN1 primary antibody (#27, AbClon), and washed (30 min) with 0.1% TX-100 in PBS.

Techniques: Knockdown, Quantitative RT-PCR, Synthesized, Western Blot, Expressing, Immunohistochemical staining, Staining, Microarray

Journal: Cancer Biology & Therapy

Article Title: Engrailed 1 overexpression as a potential prognostic marker in quintuple-negative breast cancer

doi: 10.1080/15384047.2018.1423913

Figure Lengend Snippet: EN1 expression with respect to breast cancer subtype and clinicopathological characteristics of patients with TNBC.

Article Snippet: The cells were then blocked with 3% non-fat milk in 0.1% TX-100 in PBS, incubated (overnight, 4°C) with an anti-EN1 primary antibody (#27, AbClon), and washed (30 min) with 0.1% TX-100 in PBS.

Techniques: Expressing, Significance Assay

Journal: Cancer Biology & Therapy

Article Title: Engrailed 1 overexpression as a potential prognostic marker in quintuple-negative breast cancer

doi: 10.1080/15384047.2018.1423913

Figure Lengend Snippet: EN1 protein expression is associated with poor survival in patients with quintuple-negative breast cancer (QNBC). The impact of nuclear and/or cytoplasmic EN1 protein localization on the overall survival (OS) of patients with (A) TNBC, (B) BLBC, and (C) QNBC was analyzed using the Kaplan-Meier method. (C) The impact of EN1 expression on EFS in patients with QNBC was analyzed using the Kaplan-Meier method.

Article Snippet: The cells were then blocked with 3% non-fat milk in 0.1% TX-100 in PBS, incubated (overnight, 4°C) with an anti-EN1 primary antibody (#27, AbClon), and washed (30 min) with 0.1% TX-100 in PBS.

Techniques: Expressing