Journal: Cancer Science

Article Title: Design and Evaluation of Eb 4 Mab ‐7‐ mG 2a : A Dual‐Action Anti‐ EphB4 Monoclonal Antibody for Targeted Breast Cancer Therapy

doi: 10.1111/cas.70198

Figure Lengend Snippet: Prognostic significance of EphB4 expression in breast cancer and establishment of Eb 4 Mab‐7‐mG 2a . (A) Kaplan–Meier survival analysis of OS in breast cancer patients based on the expression levels of EphB family genes (EphB1, EphB2, EphB3, EphB4, and EphB6) using RNA‐seq data from KMplot. Among these genes, high expression of EphB2 and EphB4 was significantly associated with poor prognosis (log‐rank p < 0.05). Patients were then stratified into high and low expression groups using the optimal cutoff within the interquartile range. (B) Generation of a mouse IgG 2a version of the anti‐EphB4 monoclonal antibody (Eb 4 Mab‐7‐mG 2a ). The V H and V L regions of B4Mab‐7 (mouse IgG 1 ) were cloned into expression vectors containing mouse IgG 2a C H and C L regions. These vectors were transfected into BINDS‐09 cells to produce Eb 4 Mab‐7‐mG 2a . (C) Left: Expression of EphB4 and β‐Actin in CHO‐K1, CHO/EphB4, MCF‐7, and EphB4‐KO MCF‐7 cells. Right: Expression of EphB4 and GAPDH in normal human adult breast tissues, MCF10A (non‐tumorigenic mammary epithelial cells), MCF‐7, BT‐474, SK‐BR‐3, MDA‐MB‐468, and HBC5 cells.

Article Snippet: Aside from cell lysates, a commercially available normal human breast tissue lysate (Total Protein–Human Adult Normal Tissue: Breast, Novus Biologicals, Littleton, CO, USA) was used to assess expression in normal breast tissue.

Techniques: Expressing, RNA Sequencing, Clone Assay, Transfection

Journal: Scientific Reports

Article Title: Acetylation of conserved DVL-1 lysines regulates its nuclear translocation and binding to gene promoters in triple-negative breast cancer

doi: 10.1038/s41598-019-52723-3

Figure Lengend Snippet: Dishevelled-1 proteins are highly expressed in triple-negative breast cancer cell lines. Total mRNA was isolated from different breast cancer cell lines: human non-cancer mammary epithelial cell line (MCF10A and MCF12F), hormone receptor positive breast cancer cells (MCF-7, T-47D) and triple-negative breast cancer cell lines (MDA-MB-231, BT-549 and MDA-MB-468). (A) Real-time PCR (qRT-PCR) analysis of endogenous DVL-1 gene was performed using intron-spanning primers. All results are expressed as mean ± SEM and considered significant at *p < 0.05, **p < 0.01 and ***p < 0.001. (B) The protein expression patterns of endogenous DVL-1 were analysed by Western blotting in breast cells lines, as described above, along with breast normal whole tissue lysates (NT1, NT2, and NT3). The membranes were probed with two different DVL-1 specific antibodies (D3570; Sigma and sc-8025; Santa Cruz Biotechnology, Inc), and β-actin was included as a control (see Supplementary Fig. ).

Article Snippet: Human breast whole tissue lysates (normal) were purchased from GeneTex (GTX24006), Novus Biological (NB820-59203) and G-Biosciences (NLH-13).

Techniques: Isolation, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Expressing, Western Blot, Control