Journal: European Journal of Oral Sciences

Article Title: Higher expression of WNT 5A protein in oral squamous cell carcinoma compared with dysplasia and oral mucosa with a normal appearance

doi: 10.1111/eos.12352

Figure Lengend Snippet: Expression of WNT 5A in different tissue regions. (A and B) Nuclear expression of WNT5A in oral mucosa with normal appearance. (C and D) Cytoplasmic and nuclear expression of WNT5A in severe‐grade dysplasia (black arrows). (E and F) Cancer islands; black arrows indicate expression of WNT5A in the cytoplasm in the periphery of cancer islands and red arrows indicate the absence of expression of WNT5A in the central part of the cancer islands. Scale bar = 50 μ m.

Article Snippet: The cells were cultured in six‐well plates until 80% confluence was reached, rinsed with PBS, incubated with serum‐free medium overnight, rinsed again with PBS, and treated either with 0.1% BSA in PBS as the control or with 0.4 μ g/ml of recombinant WNT5A (rWNT5A; 645‐WN; R&D Systems, Minneapolis, MN, USA) in serum‐free medium.

Techniques: Expressing

Journal: European Journal of Oral Sciences

Article Title: Higher expression of WNT 5A protein in oral squamous cell carcinoma compared with dysplasia and oral mucosa with a normal appearance

doi: 10.1111/eos.12352

Figure Lengend Snippet: Effect of recombinant WNT 5A ( rWNT 5A) on expression of β ‐catenin and E‐cadherin in SCC 9 and SCC 25. (A, E) Representative western blots of expression of active β ‐catenin, total β ‐catenin, and E‐cadherin after stimulation with rWNT 5A. (B, F) Quantification of relative E‐cadherin; (C, G) quantification of active β ‐catenin; (D, H) quantification of total β ‐catenin. All quantifications were performed on four separate experiments.

Article Snippet: The cells were cultured in six‐well plates until 80% confluence was reached, rinsed with PBS, incubated with serum‐free medium overnight, rinsed again with PBS, and treated either with 0.1% BSA in PBS as the control or with 0.4 μ g/ml of recombinant WNT5A (rWNT5A; 645‐WN; R&D Systems, Minneapolis, MN, USA) in serum‐free medium.

Techniques: Recombinant, Expressing, Western Blot

Journal: European Journal of Oral Sciences

Article Title: Higher expression of WNT 5A protein in oral squamous cell carcinoma compared with dysplasia and oral mucosa with a normal appearance

doi: 10.1111/eos.12352

Figure Lengend Snippet: Expression of WNT5A, β ‐catenin, and E‐cadherin in oral mucosa with a normal appearance, dysplasia, and at the invasive front of oral squamous cell carcinoma (OSCC)

Article Snippet: The cells were cultured in six‐well plates until 80% confluence was reached, rinsed with PBS, incubated with serum‐free medium overnight, rinsed again with PBS, and treated either with 0.1% BSA in PBS as the control or with 0.4 μ g/ml of recombinant WNT5A (rWNT5A; 645‐WN; R&D Systems, Minneapolis, MN, USA) in serum‐free medium.

Techniques: Expressing, Staining

Journal: International Journal of Nanomedicine

Article Title: Second-generation aptamer-conjugated PSMA-targeted delivery system for prostate cancer therapy

doi: 10.2147/ijn.s23747

Figure Lengend Snippet: Figure 5 Bcl-2, Cyclin D1, and Wnt3a knock down following miRNA/PAMAM-PEG-APT complexes delivery to LNCaP cells. The cells were treated with miRNA/ PAMAM-PEG-APT (miRNA) and NC-miRNA/PAMAM-PEG-APT (NC-miRNA) complexes and processed for immunoblotting with anti-Bcl-2, anti-Cyclin D1, and anti-Wnt3a antibodies at 24, 48, and 72 hours post-transfection. β-actin was a loading control. Abbreviations: PAMAM, polyamidoamine; PEG, polyethylene glycol; APT, aptamer.

Article Snippet: The blotted membrane was immunostained with antibodies specific to anti-Bcl-2 (BioWorld, Visalia, CA), anti-Cyclin D1 (BioWorld) and anti-Wnt3a antibodies (R&D Systems Inc, Minneapolis, MN).

Techniques: Knockdown, Western Blot, Transfection, Control

Journal: Cancer Research Communications

Article Title: WNT4 Regulates Cellular Metabolism via Intracellular Activity at the Mitochondria in Breast and Gynecologic Cancers

doi: 10.1158/2767-9764.CRC-23-0275

Figure Lengend Snippet: BioID supports WNT4 localization to the mitochondria. A, Proteins enriched in HT1080 Wnt-BirA versus parental HT1080 cells lacking BirA construct expression. B, Overlap of proteins identified in HT1080 (A) versus HT1080-PKO and MM134 identifies n = 72 “high-confidence” WNT4-associated proteins. C, Gene ontology analysis for cellular compartment for WNT3A- versus WNT4-associated proteins. Dashed line = 1.3 ( P = 0.05). D, Network analysis of WNT3A- versus WNT4-associated proteins via subcell barcode. Enrichments against cell line HCC287 background shown; parallel results observed with other cell line background data, for example, MCF7. E, Proteins with predicted cytosolic or mitochondrial localization (subcell barcode) among “high-confidence” WNT4-associated proteins. Red = predicted mitochondrial localization, pink = mTOR complex in mitochondrial dynamics, biogenesis, and autophagy. F, Biotin treatment and streptavidin pulldown was performed as for MS studies, and candidate WNT4-associated proteins from E detected by immunoblotting. Total protein by Ponceau.

Article Snippet: Blots were probed with Streptavidin-HRP (Cell Signaling Technology #3999; RRID:AB_10830897) or antibodies used according to manufacturer's recommendations: WNT4 (R&D Systems, MAB4751; RRID:AB_2215448); WNT3A (Novus Biologicals, MAB13242); anti-HA-HRP conjugate (Cell Signaling Technology #2999; RRID:AB_1264166); DHRS2 (Sigma HPA053915; RRID:AB_2682307); mTOR (Cell Signaling Technology #2983; RRID:AB_2105622); STAT1 (Sigma HPA000931; RRID:AB_1080100).

Techniques: Construct, Expressing, Western Blot

Journal: PLoS ONE

Article Title: A FOXM1 Dependent Mesenchymal-Epithelial Transition in Retinal Pigment Epithelium Cells

doi: 10.1371/journal.pone.0130379

Figure Lengend Snippet: A. Quantification of change in cell density (number of DAPI positive nuclei per cm 2 imaged area) upon FOXM1 overexpression or knockdown, 72h post transfection. Data is normalized to appropriate controls (Empty vector for pFOXM1 and non-targeting siRNA for siFOXM1). Bars represent Mean + SD (n = 4). P<0.0001 (Student’s t-test). B. Heatmap showing changes in gene expression of a panel of representative markers over a timecourse of RPE culture where cells are seeded at high (100000 cells/cm 2 ) or low (8000 cells/cm 2 ) density. C. Plot showing differential expression of BMP7 and Wnt5B transcripts extrapolated from the microarray data. The shaded area represents 95% confidence intervals around the point estimates (circles) of the difference between the mean high density expression vs the mean low density expression.

Article Snippet: Where required, media was supplemented with recombinant human Wnt5B (500ng/ml; R&D Systems), BMP-4/7 (75ng/ml; R&D Systems), Thiostrepton (Sigma), LDN-193189 (10μM; Stemgent), WAY-262611 (10μM; Enzo Lifesciences).

Techniques: Over Expression, Knockdown, Transfection, Plasmid Preparation, Gene Expression, Quantitative Proteomics, Microarray, Expressing

Journal: PLoS ONE

Article Title: A FOXM1 Dependent Mesenchymal-Epithelial Transition in Retinal Pigment Epithelium Cells

doi: 10.1371/journal.pone.0130379

Figure Lengend Snippet: A. Immunocytochemistry for PMEL17 where cells are seeded at either low density (16000 cells/cm 2 ) in the presence or absence of BMP4/7 (top left) or at high density (25000 cells/cm 2 ) in the presence or absence of Wnt5B (bottom left) and cultured for a period of 14 days. Also shown is the expression of BEST1 under the same conditions (top and bottom right). ACTB and B2M are used as housekeeping genes. Bars represent Mean + SD (n = 3). B. Quantification of immunocytochemistry for % CRALBP at Day 21 where cells are either treated with media alone (Control) or media supplemented with 10μM LDN-193189 added at Day 2,4,6,8,11,14 or 18. * indicates significant difference between control and compound treatment (One way ANOVA with Dunnett’s multiple comparisons). C. Quantification of immunocytochemistry for % CRALBP at Day 28 where cells are either treated with media alone (Control) or media supplemented with 10μM WAY-262611 added at Day 2,7,14 or 21. * indicates significant difference between control and compound treatment (One way ANOVA with Dunnett’s multiple comparisons). D. qPCR based measurement of BMP7 and Wnt5B transcript expression at Day 10 post siFOXM1 transfection (relative to transfection with non-targeting siRNA used as a control). GAPDH , HPRT1 and IPO8 were used as housekeeping genes. Bars represent Mean + SD (n = 3). P<0.05 (Student’s t-test).

Article Snippet: Where required, media was supplemented with recombinant human Wnt5B (500ng/ml; R&D Systems), BMP-4/7 (75ng/ml; R&D Systems), Thiostrepton (Sigma), LDN-193189 (10μM; Stemgent), WAY-262611 (10μM; Enzo Lifesciences).

Techniques: Immunocytochemistry, Cell Culture, Expressing, Control, Transfection

Journal: PLoS ONE

Article Title: A FOXM1 Dependent Mesenchymal-Epithelial Transition in Retinal Pigment Epithelium Cells

doi: 10.1371/journal.pone.0130379

Figure Lengend Snippet: RPE first acquire a mesenchymal morphology upon dissociation and culture followed by proliferation and mesenchymal-epithelial transition to re-uptake an epithelial phenotype. Proliferation of RPE is directly regulated by FOXM1 which also affects expression of BMP7 and Wnt5B by an unknown mechanism. Both these activities are required for successful MET and epithelialization.

Article Snippet: Where required, media was supplemented with recombinant human Wnt5B (500ng/ml; R&D Systems), BMP-4/7 (75ng/ml; R&D Systems), Thiostrepton (Sigma), LDN-193189 (10μM; Stemgent), WAY-262611 (10μM; Enzo Lifesciences).

Techniques: Expressing