Journal: Cellular and Molecular Bioengineering

Article Title: Multicompartmentalized Microvascularized Tumor-on-a-Chip to Study Tumor-Stroma Interactions and Drug Resistance in Ovarian Cancer

doi: 10.1007/s12195-024-00817-y

Figure Lengend Snippet: Microfluidic device mimics drug penetration gradients in OC-TME. a Schematic representation of method used for doxorubicin penetration assay, where doxorubicin (100 µM) was injected in both circulation channels and images were taken in time-lapse fluorescent microscope up to 24 h. b Representative images at 5 min, 30 min, 2, 4, 7 and 24 h of doxorubicin penetration. Scale Bar = 1 mm. c Quantification of the fluid velocity (μm/s) inside the microfluidic device over the time (h) (left) and MFI of doxorubicin penetration inside the cancer and stromal chambers at each time point (right). d Schematic representation of method used for doxorubicin uptake, cancer cells were cultured surrounded by different stromal cells (EC, NF, or CAF) or blank scaffold as a control for 5 days and then doxorubicin (100 µM) was injected in both circulation channels and images were taken at 24 h. e Representative images of doxorubicin uptake by cells at 24 h. Scale Bar = 1 mm. Figure a and d created with Biorender.com

Article Snippet: The images were taken at different time points (0; 0.25; 0.5; 0.75, 1; 1.25; 1.5; 1.75; 2, 2.5; 3; 4; 5; 6; 7; 8; 24 hours) for drug penetration and at 24 h for drug uptake using Nikon Eclipse Ni-E fluorescent microscope (EX 559.5 nm/EM 645 nm) at 10× magnification.

Techniques: Injection, Microscopy, Cell Culture, Control

Journal: Frontiers in Immunology

Article Title: Wnt5A Signaling Promotes Defense Against Bacterial Pathogens by Activating a Host Autophagy Circuit

doi: 10.3389/fimmu.2018.00679

Figure Lengend Snippet: Reduction in Wnt5A level promotes bacterial infection in mouse models. (A) Liposomal formulation of inhibitor of Wnt production (IWP2) (LI) and empty liposome (L) were administered intraperitoneally to mice prior to Pseudomonas aeruginosa (PA) challenge and mice sacrificed after 2 h ( n = 5). The difference in bacterial load [colony forming unit (CFU) recovered from peritoneal cells] between the two groups is represented in [ (A) i,ii]. [ (A) iii] depicts the inhibitory effect of LI on Wnt5A secretion as estimated from peritoneal lavage. (B) LI and L were administered by intravenous route to mice prior to intranasal inoculation with PA and mice sacrificed after 2 h ( n = 5). The difference in bacterial load is represented in [ (B) i,ii]. Inhibitory effect of LI on Wnt5A but not Wnt3A secretion after intravenous administration is demonstrated in mice sera by western blotting in [ (B) iii]. (C) IWP2 drug and vehicle control (PBS) were administered as oral gavage for five consecutive days before cecal ligation and puncture (CLP)-induced polymicrobial sepsis and mice sacrificed after 12 h ( n = 4). [ (C) i,ii] depict the difference in bacterial load in the peritoneal lavage between the IWP2 and PBS groups. [ (C) iii] depicts the inhibitory effect of IWP2 on Wnt5A secretion as estimated from peritoneal lavage and serum. (D) PA peritonitis was induced in Wnt5A +/+ and Wnt5A +/− mice and mice sacrificed after 2 h of infection ( n = 9). The difference in bacterial load between the two groups is presented in [ (D) i,ii]. Peritoneal cells of Wnt5A +/− mice express less Wnt5A than the corresponding controls [ (D) iii]. (E) Panels (i,ii) depict the difference in the bacterial load harbored by peritoneal macrophages in LI vs. L and Wnt5A +/− vs. Wnt5A +/+ sets of mice. Dots indicated by arrows in insets represent the propidium iodide stained bacteria in the macrophages. Data represented as mean ± SEM; * p ≤ 0.05, ** p ≤ 0.005, *** p ≤ 0.0005.

Article Snippet: Recombinant mouse Wnt5A (645-WN), anti-mouse Wnt5A antibody (AF-645), and anti-human Wnt5A antibody (MAB-645) were purchased from R&D Systems.

Techniques: Infection, Formulation, Western Blot, Control, Ligation, Staining, Bacteria

Journal: Frontiers in Immunology

Article Title: Wnt5A Signaling Promotes Defense Against Bacterial Pathogens by Activating a Host Autophagy Circuit

doi: 10.3389/fimmu.2018.00679

Figure Lengend Snippet: Pseudomonas aeruginosa (PA) and Streptococcus pneumoniae (SP) infection lead to reduction of both cell associated and secreted Wnt5A. There is decrease in both cell associated (A–C) and secreted (D) Wnt5A upon infection with PA (strain: PA14 and PA01) and SP (strain: A66) using different multiplicity of infection (MOI) ( n = 3). MOI 10 infection of PA for different lengths of time produces similar results (E,F) ( n = 3). (G) Wnt5A level in sputum samples of COPD patients with and without PA infection ( n = 5). Data represented as mean ± SEM; * p ≤ 0.05, ** p ≤ 0.005, *** p ≤ 0.0005.

Article Snippet: Recombinant mouse Wnt5A (645-WN), anti-mouse Wnt5A antibody (AF-645), and anti-human Wnt5A antibody (MAB-645) were purchased from R&D Systems.

Techniques: Infection

Journal: Frontiers in Immunology

Article Title: Wnt5A Signaling Promotes Defense Against Bacterial Pathogens by Activating a Host Autophagy Circuit

doi: 10.3389/fimmu.2018.00679

Figure Lengend Snippet: Wnt5A signaling promotes internalization and killing of pathogenic bacteria. (A) Reverse transcription PCR analysis (upper panel) and immunoblot (lower panel) demonstrating Fz5 (putative Wnt5A receptor) expression in RAW 264.7 macrophages using anti-Frizzled-5 antiserum (Immune serum) and pre-immune serum (control). (B–D) Effect of rWnt5A on internalization and total killing of Streptococcus pneumoniae (SP) (B) , Pseudomonas aeruginosa (PA): PA14 (C) , and PA01 (D) as estimated by colony forming unit (CFU) ( n = 3). (E–G) Facilitated intracellular killing of SP (E) , PA14 (F) , and PA01 (G) in RAW 264.7 as estimated by percent CFU recovered at different time points (T1–T4) after 1 h infection (T0) ( n = 3). (H) Wnt5A-mediated killing of PA in mouse peritoneal macrophages ( n = 3). (I,J) Wnt5A-induced intracellular killing of PA14 (I) and SP (J) in J774A.1 at different time points (T1–T4) after 1 h infection (T0) ( n = 3). (K) Similar killing activity of Wnt5A on RAW 264.7 macrophages infected with PA strain isolated from COPD patient sputum ( n = 3). (L) rWnt5A facilitated bacterial killing depicted by confocal microscopy. Red dots indicate propidium iodide stained bacteria 3 h postinfection in RAW 264.7 macrophages pretreated either with rWnt5A or PBS. (M,N) Inhibitor of Wnt production (IWP2)-mediated inhibition (24 h of IWP2 treatment after 1 h infection) of Wnt5A production leading to significant sustenance of SP (M) and PA (N) in RAW 264.7 ( n = 6). (O) ELISA documenting IWP2 mediated reduction in Wnt5A secretion from RAW 264.7 macrophages ( n = 3). Data represented as mean ± SEM; * p ≤ 0.05, ** p ≤ 0.005, *** p ≤ 0.0005.

Article Snippet: Recombinant mouse Wnt5A (645-WN), anti-mouse Wnt5A antibody (AF-645), and anti-human Wnt5A antibody (MAB-645) were purchased from R&D Systems.

Techniques: Bacteria, Reverse Transcription, Western Blot, Expressing, Control, Infection, Activity Assay, Isolation, Confocal Microscopy, Staining, Inhibition, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Immunology

Article Title: Wnt5A Signaling Promotes Defense Against Bacterial Pathogens by Activating a Host Autophagy Circuit

doi: 10.3389/fimmu.2018.00679

Figure Lengend Snippet: Wnt5A-mediated bacterial killing is Rac1–Dvl dependent and involves actin assembly. rWnt5A-induced clearance of bacteria [ Streptococcus pneumoniae (SP) and Pseudomonas aeruginosa (PA)] was inhibited by Rac1 inhibitor (Rac1i) (A,B) and Dvl inhibitor (Dvli) (C,D) at 3 h (T3) postinfection ( n = 3). PBS and DMSO act as vehicle control. (E) Inhibitory effect of Rac1i and Dvli on Wnt5A-induced actin polymerization as detected by phalloidin staining. (F) Difference in phalloidin fluorescence intensities calculated by NIH-ImageJ software ( n = 3). (G,H) Inhibitory effect of cytochalasin D (CytD) (postinfection) on Wnt5A-mediated bacterial killing. Data represented as mean ± SEM; * p ≤ 0.05, ** p ≤ 0.005, *** p ≤ 0.0005.

Article Snippet: Recombinant mouse Wnt5A (645-WN), anti-mouse Wnt5A antibody (AF-645), and anti-human Wnt5A antibody (MAB-645) were purchased from R&D Systems.

Techniques: Bacteria, Control, Staining, Fluorescence, Software

Journal: Frontiers in Immunology

Article Title: Wnt5A Signaling Promotes Defense Against Bacterial Pathogens by Activating a Host Autophagy Circuit

doi: 10.3389/fimmu.2018.00679

Figure Lengend Snippet: Wnt5A-induced bacterial clearance is mediated through autophagy. (A–D) Effect of 3-methyladenine (3-MA, 1 mM) and wortmanin (WM, 100 nM), respectively, on rWnt5A-mediated killing of Pseudomonas aeruginosa (PA) and Streptococcus pneumoniae (SP) ( n = 3). Percent killing was calculated for 3 h (T3) killing time-point. DMSO (DM) and PBS were used as vehicle control. (E) LC3BII accumulation assessed at different killing time-points (T0–T4) for rWnt5A and PBS pretreated RAW 264.7 cells infected separately with PA and SP by western blotting ( n = 3). T0 represents bacterial killing in 0 time (post 1 h of infection). (F) Influence of WM, 3-MA, Rac1 inhibitor, and Dvl inhibitor (Dvli) on rWnt5A-induced LC3BII accumulation at 3 h (T3) postinfection with PA ( n = 3). (G,H) LC3-punctae in Wnt5A vs. PBS sets as assessed by confocal microscopy (G) and punctae dot calculation (H) ( n = 3). (I) Presence of Wnt5A in L5A conditioned medium (L5A: L cells expressing Wnt5A) but not in L conditioned medium (used as control) ( n = 3). (J,K) Difference in bacterial load between L5A Phg (phagosome isolated from L5A treated cells 2 h postinfection: T2) and L Phg (corresponding control) ( n = 3). (L) Higher levels of LC3BII, ATG5-ATG12, Rac1, p62, LAMP-1, and p-Dvl-2 (phosphorylated Disheveld-2) in L5A Phg compared to L Phg. Both sets contain similar levels of precursor and mature forms of cathepsin D (CathD). Data represented as mean ± SEM; * p ≤ 0.05, ** p ≤ 0.005, *** p ≤ 0.0005.

Article Snippet: Recombinant mouse Wnt5A (645-WN), anti-mouse Wnt5A antibody (AF-645), and anti-human Wnt5A antibody (MAB-645) were purchased from R&D Systems.

Techniques: Control, Infection, Western Blot, Confocal Microscopy, Expressing, Isolation

Journal: Frontiers in Immunology

Article Title: Wnt5A Signaling Promotes Defense Against Bacterial Pathogens by Activating a Host Autophagy Circuit

doi: 10.3389/fimmu.2018.00679

Figure Lengend Snippet: Ulk1 kinase activity (KA) is needed for rWnt5A-mediated bacterial clearance. (A) At 2 h killing time point (T2), immunoprecipitated (IP) Ulk1 from rWnt5A-treated RAW 264.7 cells demonstrates greater Ulk1 KA than the corresponding control as judged by phosphorylation of myelin basic protein (MBP). Significant difference in KA not noted in uninfected cells pretreated with Wnt5A or PBS ( n = 3). Inp (Input) is same in all cases. (B) rWnt5A-mediated killing inhibited by Ulk1 kinase inhibitor (Ulk1i) at 3 h (T3) killing time-point ( n = 3). DM (DMSO) and PBS were used as vehicle control. (C) rWnt5A-mediated LC3BII accumulation inhibited in presence of Ulk1i ( n = 3). Data represented as mean ± SEM; *** p ≤ 0.0005.

Article Snippet: Recombinant mouse Wnt5A (645-WN), anti-mouse Wnt5A antibody (AF-645), and anti-human Wnt5A antibody (MAB-645) were purchased from R&D Systems.

Techniques: Activity Assay, Immunoprecipitation, Control, Phospho-proteomics

Journal: Frontiers in Immunology

Article Title: Wnt5A Signaling Promotes Defense Against Bacterial Pathogens by Activating a Host Autophagy Circuit

doi: 10.3389/fimmu.2018.00679

Figure Lengend Snippet: Transmission electron microscopy (TEM) demonstrating Wnt5A-induced autophagosome like moieties and bacterial killing after PA infection. Lesser number of bacteria present in Wnt5A-treated cells compared to PBS-treated cells (A,B) . Arrows point to double membrane or multilamellar structures encapsulating bacteria in Wnt5A-treated macrophages (C–F) . M represents mitochondria. D denotes degraded bacteria.

Article Snippet: Recombinant mouse Wnt5A (645-WN), anti-mouse Wnt5A antibody (AF-645), and anti-human Wnt5A antibody (MAB-645) were purchased from R&D Systems.

Techniques: Transmission Assay, Electron Microscopy, Infection, Bacteria, Membrane

Journal: Cells

Article Title: Wnt5A and TGFβ1 Converges through YAP1 Activity and Integrin Alpha v Up-Regulation Promoting Epithelial to Mesenchymal Transition in Ovarian Cancer Cells and Mesothelial Cell Activation

doi: 10.3390/cells11020237

Figure Lengend Snippet: Higher expression levels of TGFβ components, Wnt5A, and Hippo-related genes in HGSOC specimens. ( A ) The heatmap shows the expression levels of TGFβ components, Wnt5A, and Hippo-related genes in Serous human EOC specimens (HG: high grade; LG: low grade; BL: borderline) and normal ovaries obtained by hierarchical cluster analysis. Each column in the figure represents a sample, and each row represents a gene. The colors in the graph indicate the magnitude of gene expression in the sample. The black–red gradient indicates that the genes are medium-high, and the blue indicates low gene expression. ( B ) The box plot shows differential gene expression levels in tumor specimens compared to normal ovaries. ( C ) The correlation analysis between TGFβ components, Wnt5A, and Hippo-related genes. ( D ) The protein–protein interaction (PPI) network of Wnt5A and TGFβ signaling components showed the interaction between the molecules involved in these signaling pathways; the thickness of the edge indicates a strong interaction between the two proteins. PPI enrichement p value = 1 −16 . ( E ) The Venn diagram shows the common pathways between TGFβ and Wnt5A signaling.3.2. Wnt5A Modulates Smad2/3 Activation in OvCa Cells.

Article Snippet: Recombinant human Wnt5A was obtained from R&D Systems (Minneapolis, MN, USA).

Techniques: Expressing, Gene Expression, Protein-Protein interactions, Activation Assay

Journal: Cells

Article Title: Wnt5A and TGFβ1 Converges through YAP1 Activity and Integrin Alpha v Up-Regulation Promoting Epithelial to Mesenchymal Transition in Ovarian Cancer Cells and Mesothelial Cell Activation

doi: 10.3390/cells11020237

Figure Lengend Snippet: Wnt5A modulates Smad2/3 activation in ovarian cancer cells. The SKOV-3, OVCAR-3, and CAOV-4 cells were transfected with siRNA Wnt5A or Scrambled siRNA (Scr). ( A ) The expression level of Wnt5A, TGFβ1, pSmad2/3 was assessed by immunoblotting in multicellular aggregates (MCAs) (Left panel). GAPDH levels were used as an internal control. The right panel shows the quantification of bands from three independent experiments ( B ) Immunolocalization of pSmad2/3 in Wnt5A silenced cells in the absence or presence of rhTGFβ1 (10 ng/mL) for 1 h (left panel). The right panel shows the percent of pSmad2/3 positive cells. Original magnification, ×400. *: p < 0.05, **: p < 0.01 and ***: p < 0.001 compared to scramble (Scr), n = 3.

Article Snippet: Recombinant human Wnt5A was obtained from R&D Systems (Minneapolis, MN, USA).

Techniques: Activation Assay, Transfection, Expressing, Western Blot, Control

Journal: Cells

Article Title: Wnt5A and TGFβ1 Converges through YAP1 Activity and Integrin Alpha v Up-Regulation Promoting Epithelial to Mesenchymal Transition in Ovarian Cancer Cells and Mesothelial Cell Activation

doi: 10.3390/cells11020237

Figure Lengend Snippet: Wnt5A is required for TGFβ1-induced migration and invasion of ovarian cancer cells. ( A ) Cell migration and ( B ) Cell invasion siRNA Wnt5A or siRNA scrambled transfected cells (Scr) were seeded at 2.5 × 10 4 cell density on the upper chamber of transwells in the presence or absence of rhTGFβ1 (10 ng/mL) or rhWnt5A (600 ng/mL) in serum-free medium for 14 h. Cell invasion was performed using matrigel-coated transwells. Photos represent one of the three independent experiments. ( C ) Migrated and invaded cells were quantified, and the results were expressed as mean ± SD, n = 3. Original magnification,×100. *: p < 0.05, **: p < 0.01 compared to untreated control cells (Ctrl) or Scr.

Article Snippet: Recombinant human Wnt5A was obtained from R&D Systems (Minneapolis, MN, USA).

Techniques: Migration, Transfection, Control

Journal: Cells

Article Title: Wnt5A and TGFβ1 Converges through YAP1 Activity and Integrin Alpha v Up-Regulation Promoting Epithelial to Mesenchymal Transition in Ovarian Cancer Cells and Mesothelial Cell Activation

doi: 10.3390/cells11020237

Figure Lengend Snippet: The positive feedback loop between Wnt5A and YAP1 and inhibition of YAP1 transcriptional activity decreases Smad2/3 activation in OvCa cells. SKOV-3, OVCAR-3, and CAOV-4 cells were transfected with siRNA scrambled (Scr) or siRNA Wnt5A. ( A ) The upper panel shows YAP1 expression levels in multicellular aggregates (MCAs) of OvCa cells. The lower panels show the quantification of bands from three independent experiments. ( B ) The left panel shows the expression level of Wnt5A in the cells treated with VP (5 μM), and the right panel shows the quantification of bands from three independent experiments. ( C ) Cells as MCAs were treated as follows: rhTGFβ1 (10 ng/mL) for 1 h, rhWnt5A (600 ng/mL) for 14 h, VP (5 μM) for 1 h, VP-pretreated + rhWnt5A, and VP-pretreated + rhTGFβ1. The upper panel represents immunoblots, and the lower panels show the quantification of bands from three independent experiments. GAPDH levels were used as internal control, and results are expressed as mean ± SD. *: p < 0.05, **: p < 0.01 and ***: p < 0.001 compared to untreated control cells (Ctrl) or Scr, n = 3.

Article Snippet: Recombinant human Wnt5A was obtained from R&D Systems (Minneapolis, MN, USA).

Techniques: Inhibition, Activity Assay, Activation Assay, Transfection, Expressing, Western Blot, Control

Journal: Cells

Article Title: Wnt5A and TGFβ1 Converges through YAP1 Activity and Integrin Alpha v Up-Regulation Promoting Epithelial to Mesenchymal Transition in Ovarian Cancer Cells and Mesothelial Cell Activation

doi: 10.3390/cells11020237

Figure Lengend Snippet: The pSmad2/3 nuclear localization is reduced in verteporfin-treated cells reverted by the presence of exogenous Wnt5A. ( A ) SKOV-3, ( B ) OVCAR-3 and ( C ) CAOV-4 Cells were treated with rhWnt5A (600 ng/mL) or Verteporfin (VP) (5 μM) alone or VP-pre-treated + rhWnt5A and subcellular localization of pSmad2/3 and YAP1. The right panels shows the percent of pSmad2/3 and YAP1 positive cells. Original magnification, ×400. *: p < 0.05, **: p < 0.01 and ***: p < 0.001 compared to Ctrl, n = 3. Images were visualized by Zeiss inverted fluorescence microscopy.

Article Snippet: Recombinant human Wnt5A was obtained from R&D Systems (Minneapolis, MN, USA).

Techniques: Fluorescence, Microscopy

Journal: Cells

Article Title: Wnt5A and TGFβ1 Converges through YAP1 Activity and Integrin Alpha v Up-Regulation Promoting Epithelial to Mesenchymal Transition in Ovarian Cancer Cells and Mesothelial Cell Activation

doi: 10.3390/cells11020237

Figure Lengend Snippet: YAP1 regulates Wnt5A-induced integrin av and Smad2/3 activation. SKOV-3, OVCAR-3, and CAOV-4 cells were transfected with siRNA Scrambled (Scr) or Wnt5A. ( A ) Integrin αv expression level was assessed by immunoblotting in multicellular aggregates (MCAs) OvCa cells (upper panel) and quantifying bands from three experiments in the lower panel. ( B ) The cells were pre-treated with VP (5 μM) 1 h or treated with rhWnt5A (600 ng/mL) for 14 h alone or VP-pretreated + rhWnt5A then expression levels of integrin αv was determined in MCAs OvCa cells (Left panel), and quantification of bands (right panel). ( C ） The Wnt5A overexpressing OVCAR-3 (C3/OVCAR-3) and SKOV-3 clones (C9/SKOV-3) were treated with CWHM-12 (10 μM) for 24 h. Immunoblot of TGFβ1, pSmad2/3, YAP1 determined in MCAs OvCa cells (Left panel) and quantification of bands (right panels). The lower panel shows the quantification of bands from three independent experiments. GAPDH levels were used as an internal control, and results are expressed as mean ± SD. (D) The immunolocalization of pSmad2/3 and YAP-1 in CWHM-12-treated cells compared to control. Original magnification, ×400. *: p < 0.05, **: p < 0.01 and ***: p < 0.001 compared to untreated control cells (Ctrl) or Scr.

Article Snippet: Recombinant human Wnt5A was obtained from R&D Systems (Minneapolis, MN, USA).

Techniques: Activation Assay, Transfection, Expressing, Western Blot, Clone Assay, Control

Journal: Cells

Article Title: Wnt5A and TGFβ1 Converges through YAP1 Activity and Integrin Alpha v Up-Regulation Promoting Epithelial to Mesenchymal Transition in Ovarian Cancer Cells and Mesothelial Cell Activation

doi: 10.3390/cells11020237

Figure Lengend Snippet: Wnt5A enhances mesothelial cell activation through Smad2/3 and YAP1 activation. The human primary omental mesothelial cells (HPOMCSs) were treated with rhWnt5A or conditioned medium (C.M) isolated from C3/OVCAR-3 clone (C3/OVCAR-3-derivedC.M), or C.M from Wnt5A silenced OVCAR-3 cells. ( A ) Immunolocalization of α-SMA, ( B ) pSmad2/3, and ( C ) YAP1. The right panels shows the percent of α-SMA, pSmad2/3, or YAP1 positive cells. Original magnification, ×400. *: p < 0.05, **: p < 0.01 and ***: p < 0.001 compared to Ctrl, n = 3.

Article Snippet: Recombinant human Wnt5A was obtained from R&D Systems (Minneapolis, MN, USA).

Techniques: Activation Assay, Isolation

Journal: Cells

Article Title: Wnt5A and TGFβ1 Converges through YAP1 Activity and Integrin Alpha v Up-Regulation Promoting Epithelial to Mesenchymal Transition in Ovarian Cancer Cells and Mesothelial Cell Activation

doi: 10.3390/cells11020237

Figure Lengend Snippet: Wnt5A enhanced mesothelial cell clearance through Smad2/3 and YAP1 activation. The following OVCAR-3 spheroids were labeled with CellTracker™ CM-DiI Dye: OVCAR-3, Wnt5A overexpressing C3/OVCAR-3 spheroids, Wnt5A silenced OVCAR-3, VP-treated OVCAR-3, and CWHM-12-treated OVCAR-3 cells. Then added to the top of the HPOMCSs monolayer. ( A ) Disaggregation and invasion of multicellular aggregates (MCAs) through HPOMCSs were followed for 24, 48, and 72 h compared to the initial time set as 2 h. Phase-contrast photos after 72 h showed the mesothelial cell retraction. ( B ) The surface area of spheroids was calculated as described in material and methods. The results are expressed as mean + SD of at least three independent experiments. **: p < 0.01 compared to untreated OVCAR-3 spheroids (Ctrl). Original magnification: ×100.

Article Snippet: Recombinant human Wnt5A was obtained from R&D Systems (Minneapolis, MN, USA).

Techniques: Activation Assay, Labeling

Journal: Cells

Article Title: Wnt5A and TGFβ1 Converges through YAP1 Activity and Integrin Alpha v Up-Regulation Promoting Epithelial to Mesenchymal Transition in Ovarian Cancer Cells and Mesothelial Cell Activation

doi: 10.3390/cells11020237

Figure Lengend Snippet: Model of Wnt5A involvement in EMT and mesothelial activation and clearance. Wnt5A derived from ovarian cancer cells through cytoskeletal rearrangement could directly or indirectly cause YAP1 phosphorylation, which translocates into the nucleus and induces TGFβ1, integrin αv, Wnt5A, and other EMT markers in this drawing. Integrin αv, in turn, may activate extracellular latent-TGFβ1 and play a pivotal role in the EMT process. In addition, YAP1 may cause retention of Smad2/3 in the nucleus, thereby prolonging their biological activity.

Article Snippet: Recombinant human Wnt5A was obtained from R&D Systems (Minneapolis, MN, USA).

Techniques: Activation Assay, Derivative Assay, Phospho-proteomics, Activity Assay