Journal: PLoS Genetics

Article Title: The planar cell polarity protein VANG-1/Vangl negatively regulates Wnt/β-catenin signaling through a Dvl dependent mechanism

doi: 10.1371/journal.pgen.1007840

Figure Lengend Snippet: ( A ) TOPFLASH reporter activity in wild type and Vangl1/2 double mutant (DKO) HEK293 cells treated with 100 ng/ml Wnt3a or the corresponding amount of 0.1% BSA in PBS in the absence (left panel) or presence (right panel) of R-Spondin. Rescue experiments were performed with expression of wild type and C-terminally truncated Vangl2 (fused to EGFP). Statistical significance was calculated using a Student’s t-test. *, p<0.05; **, p<0.01. ( B ) Western blot analysis of Vangl1 and Vangl2 expression in control, Vangl1/2 DKO and DKO cells expressing wild type or C-terminally truncated Vangl2 (fused to EGFP). Note that endogenous Vangl2 expression is not visible at this exposure. A longer exposure is provided in .

Article Snippet: Treatments used were: recombinant human Wnt3a (CF 5036-WN-CF RnD Systems), recombinant human R-Spondin-1 (120–38, PeproTech) and LGK 974 (974–02, Stem RD) for inhibition of endogenous Wnt ligands secretion.

Techniques: Activity Assay, Mutagenesis, Expressing, Western Blot, Control

Journal: Stem cells (Dayton, Ohio)

Article Title: IFATS collection: Adipose stromal cell differentiation is reduced by endothelial cell contact and paracrine communication: role of canonical Wnt signaling.

doi: 10.1634/stemcells.2008-0277

Figure Lengend Snippet: Figure 5. Effect of canonical Wnt signaling on adipogenic differentiation of adipose stromal cells (ASC). (A): ASC were treated with either Wnt signaling agonist (1 M; Calbiochem) or recombinant Wnt protein (Wnt3a; 10 ng/ml) for 3 days prior to anal- ysis of activation of canonical Wnt signaling using the TCF7-driven Topflash reporter as- say. Note a 60% increase in Topflash activ- ity with Wnt agonist stimulation. (B): Adi- pogenic differentiation of ASC after 3 days using gene expression analysis of PPAR and LPL by reverse transcription polymerase chain reaction. A significant (, p .03) decrease was noted in adipogenic differenti- ation upon stimulation with Wnt agonist or a Wnt3a recombinant protein. Abbreviations: BM, basal medium; DM, differentiation me- dium; PPAR, peroxisome proliferator-acti- vated receptor.

Article Snippet: Thus, we examined the specific effect of canonical Wnt signaling stimulation by either the Wnt agonist ([(2-amino-4-(3,4-(methylenedioxy)benzylamino)-6-(3-methoxyphenyl) pyrimidine)]; Calbiochem, San Diego, http://www.emdbiosciences.com) or recombinant Wnt3a protein (R&D Systems Inc., Minneapolis, http://www.rndsystems.com) on both downstream Wnt pathway activation and features of adipogenesis.

Techniques: Recombinant, Activation Assay, Gene Expression, Reverse Transcription, Polymerase Chain Reaction

Journal: Oncotarget

Article Title: FOXM1 confers to epithelial-mesenchymal transition, stemness and chemoresistance in epithelial ovarian carcinoma cells

doi:

Figure Lengend Snippet: (A) Western blots of FOXM1 and the internal control β-ACTIN in whole cell lysates of A2780CP70 cells treated for 2 days with different concentrations of WNT3A and SFRP5 peptides. (B) Ad293 cells were transfected with the indicated concentrations of pcDNA3.1 (control vector) or pcDNA3.1 encoding SFRP5 and 500 ng pLuc-FOXM1 (FOXM1 promoter luciferase vector). The cells were harvested two days after transfection, and dual luciferase reporter assays were performed. FOXM1 promoter activity was expressed as the fold change relative to the control (pcDNA3.1). Each bars represents mean ± standard error of the mean from three independent experiments. **: P < 0.01; ***: P < 0.001. (C and D) Western blots of SFRP5, FOXM1, and the internal control β-ACTIN in Ad293 cells overexpressing a control vector (Vector) or shSFRP5 vector (shSFRP5) (C), and ES-2 and TOV-21G cells (D).

Article Snippet: Recombinant human SFRP5 and WNT3a peptides were purchased from R&D Systems.

Techniques: Western Blot, Control, Transfection, Plasmid Preparation, Luciferase, Activity Assay