Journal: The Journal of Clinical Investigation

Article Title: SIX1 induces lymphangiogenesis and metastasis via upregulation of VEGF-C in mouse models of breast cancer

doi: 10.1172/JCI59858

Figure Lengend Snippet: (A) Microarray analysis demonstrates that SIX1 overexpression leads to increased VEGFC mRNA in both MCF7-SIX1 cells and MCF7-SIX1 tumors. (B) Quantitation of VEGFC gene expression in MCF7-Ctrl and MCF7-SIX1 using real-time PCR. (C) SIX1 induces VEGFC promoter activity. VEGFC promoter-luciferase reporter constructs were transiently transfected into MCF7 cells, along with increasing amounts of SIX1 and a constant amount of the SIX1 cofactor, EYA2. Luciferase activity was analyzed after 48 hours. (D) ChIP was performed to detect SIX1 presence on the VEGFC promoter in MCF7 cells transfected with SIX1 and EYA2. Protein-DNA complexes were precipitated with a SIX1-specific antibody as well as a control rabbit IgG antibody, after which real-time PCR was performed with primers that flank 5 predicted SIX1 binding sites within the VEGFC promoter (red circles denote the TGATAC binding sites; green triangles denote the ATCCTGA binding sites) as well as 1 upstream region with no predicted SIX1 binding site as a negative control. Dashed line indicates the background non-specific binding of SIX1 and x axis units are base pairs upstream of transcription start site. (E) Functional VEGF-C secreted by MCF7-Ctrl and MCF7-SIX1 cells was measured by ELISA and Western blot analysis (3 clonal isolates of MCF7-Ctrl cells [lanes 1–3] and 3 clonal isolates of MCF7-SIX1 cells [lanes 4–6]) in conditioned medium after serum starvation for 48 hours. (F) MCF7-SIX1 tumors express higher levels of VEGF-C in vivo compared with the MCF7-Ctrl tumors. Immunostaining was used to detect VEGF-C and VEGF-D. Original magnification, ×400. *P < 0.05; **P < 0.01.

Article Snippet: Using Oncomine, expression values for SIX1 and VEGFC were retrieved from the Adai microarray data set (Genentech), and the expression data were plotted to perform a correlation analysis between SIX1 and VEGFC in breast cancer cell lines representing different histologic subtypes of breast cancer (Figure A).

Techniques: Microarray, Over Expression, Quantitation Assay, Gene Expression, Real-time Polymerase Chain Reaction, Activity Assay, Luciferase, Construct, Transfection, Control, Binding Assay, Negative Control, Functional Assay, Enzyme-linked Immunosorbent Assay, Western Blot, In Vivo, Immunostaining

Journal: The Journal of Clinical Investigation

Article Title: SIX1 induces lymphangiogenesis and metastasis via upregulation of VEGF-C in mouse models of breast cancer

doi: 10.1172/JCI59858

Figure Lengend Snippet: (A) SIX1 and VEGFC expression values were retrieved from an Oncomine microarray data set (as indicated in the figure) and were plotted according to different types of cell lines or by expression value. X and y axes of the right panel indicate mRNA expression analyzed on Affymetrix U133 Plus 2.0 microarrays. (B) Representative positive and negative staining of SIX1 and VEGF-C on human breast cancer tissue sections. Original magnification, ×100. (C) Model depicting the mechanism by which SIX1 promotes metastatic dissemination. Overexpression of SIX1 leads to increased VEGF-C and stimulates lymphangiogenesis, allowing for increased escape of tumor cells through the lymphatics and increased distant metastasis. However, SIX1 is also able to augment the later stages of metastasis of cancer cells that have traveled through the vasculature, thus contributing to metastatic spread via multiple mechanisms. An extension of this finding is that while inhibitors of the VEGF-C/VEGFR3 axis may prevent lymphatic spread, inhibitors of SIX1 are expected to inhibit metastasis at multiple stages, serving as powerful antimetastatic agents.

Article Snippet: Using Oncomine, expression values for SIX1 and VEGFC were retrieved from the Adai microarray data set (Genentech), and the expression data were plotted to perform a correlation analysis between SIX1 and VEGFC in breast cancer cell lines representing different histologic subtypes of breast cancer (Figure A).

Techniques: Expressing, Microarray, Negative Staining, Over Expression

Journal: BMC Genomics

Article Title: Genome-wide characterization and expression profiling of PDI family gene reveals function as abiotic and biotic stress tolerance in Chinese cabbage ( Brassica rapa ssp . pekinensis )

doi: 10.1186/s12864-017-4277-2

Figure Lengend Snippet: Heat map showing microarray expression of 32 B. rapa PDI genes against cold and freezing treatments in two inbred lines Chiifu and Kenshin

Article Snippet: We analyzed 32 BrPDI genes expression pattern based on the microarray data set in two contrasting Chinese cabbage inbred lines, ‘Chiifu’ and ‘Kenshin’ exposed to cold and freezing stress (4 °C, 0 °C, −2 °C, and −4 °C) [ ].

Techniques: Microarray, Expressing

Journal: Frontiers in Plant Science

Article Title: A Genome-Wide Analysis Reveals Stress and Hormone Responsive Patterns of TIFY Family Genes in Brassica rapa

doi: 10.3389/fpls.2016.00936

Figure Lengend Snippet: Microarray expression represented as heat map and hierarchical clustering of 36 TIFY genes against different low temperatures such as control (C1 and K1), 4°C (C2 and K2), 0°C (C3 and K3), −2°C (C4 and K4), and −4°C (C5 and K5) in B. rapa . Here, “C” and “K” stand for “Chiifu” and “Kenshin” two inbred lines of B. rapa , respectively. Expression clusters are shown in the left (Cl1–Cl5) and gene name against each expression has mentioned at right side. Color bars with values at the base represent differential expression in microarray.

Article Snippet: Using a low temperature-treated whole-genome microarray data set, most of the JAZ genes were found to have variable transcript abundance between the contrasting inbred lines Chiifu and Kenshin of B. rapa .

Techniques: Microarray, Expressing, Control, Quantitative Proteomics