Journal: eLife

Article Title: Selective YAP activation in Procr cells is essential for ovarian stem/progenitor expansion and epithelium repair

doi: 10.7554/eLife.75449

Figure Lengend Snippet: ( A ) Illustration of superovulation strategy. Four-week-old mice were administrated with pregnant mare serum gonadotropin (PMSG), following by human chorionic gonadotropin (HCG) 2 days later. The ovaries were harvested 0.5 day after HCG injection (ovulation). ( B ) Confocal images showed abundant E-cad expression in the OSE of nonrupture sites. Scale bar, 100 μm for zoom out and 10 μm for zoom in. n = 3 mice and more than 15 images. ( C-D ) Confocal images showed lower levels of α-catenin and ZO-1 in the OSE of proximal regions surrounding the rupture sites (view #1 in C) compared with those in distal regions (view #2 in C) and nonrupture regions ( D ). Scale bar, 100 μm for zoom out and 10 μm for zoom in. n = 3 mice and 15 images.

Article Snippet: Antibody , anti-a-E-catenin (Rabbit polyclonal) , Proteintech , Cat# 12831-1-AP; RRID: AB_2087822 , IHC (1:200).

Techniques: Injection, Expressing

Journal: eLife

Article Title: Selective YAP activation in Procr cells is essential for ovarian stem/progenitor expansion and epithelium repair

doi: 10.7554/eLife.75449

Figure Lengend Snippet:

Article Snippet: Antibody , anti-a-E-catenin (Rabbit polyclonal) , Proteintech , Cat# 12831-1-AP; RRID: AB_2087822 , IHC (1:200).

Techniques: Recombinant, Plasmid Preparation, shRNA, Mutagenesis, Sequencing, Imaging, Reporter Assay, Multiplex Assay, SYBR Green Assay, Software

Journal: International Journal of Molecular Sciences

Article Title: Histological and Immunohistochemical Studies to Determine the Mechanism of Cleft Palate Induction after Palatal Fusion in Mice Exposed to TCDD

doi: 10.3390/ijms23042069

Figure Lengend Snippet: Staining images of β-catenin and α-catenin in the palatal frontal sections of control and TCDD-treated groups (scale bar: 100 µm). In the control group, β-catenin and α-catenin were positive for epithelial cells in the anterior and posterior palate, the nasal and oral mucosa, and the epithelial cord ( A , B , E , F ). In the TCDD group, β-catenin and α-catenin were positive for epithelial cells in the nasolacrimal and oral mucosa at the anterior palatal fusion but negative in the median nasolacrimal mucosa ( C , D ). β-catenin and α-catenin were positive for epithelial cells in the epithelial cord of the midline palate ( C , D , arrowheads). In the posterior palatal detachment, β-catenin and α-catenin were positive for epithelial cells in the nasal and oral mucosa ( G , H ) and in the epithelial cord near the palatal detachment ( G , H , arrowheads).

Article Snippet: Rabbit monoclonal anti-β-catenin antibody (ab32572, Abcam) and mouse monoclonal anti-α-catenin antibody (66221-1-lg, Proteintech, Rosemont, IL, USA) were used as primary antibodies to observe interepithelial cell adhesion factors from the anterior to posterior palate.

Techniques: Staining

Journal: bioRxiv

Article Title: A myosin chaperone, UNC-45A, is a novel regulator of intestinal epithelial barrier integrity and repair

doi: 10.1101/2021.11.10.467974

Figure Lengend Snippet: (A,B) Viscoelastic properties, epithelial tension (A) and viscosity (B) measurements in control and UNC-45A-depleted HT-29cf8 cell monolayers by noncontact frequency modulation atomic force microscopy (FM-AFM). Means ± SD; number of cells for control sgRNA n=35, UNC-45A sgRNA1 n=31, and UNC-45A sgRNA2 n=26; **P< 0.005; ****P< 0.0001, as compared to the control sgRNA group. (C,D) Immunofluorescence labeling of control and UNC-45A-depleted HT-29cf8 cell monolayers with antibodies recognizing either total α-catenin, or an open conformation of α-catenin (α18-α-catenin antibody). Representative confocal images (C) and quantification of the labeling intensity (D) is shown. Each dot represents an averaged signal intensity of 200 cell-cell contacts measured in four different images. Means ± SE (n=4); **P< 0.005, as compared to the control sgRNA group. Scale bar, 20 μm. (E,F) Contraction of collaged I gel with embedded control or UNC-45A-depleted HT-29cf8 cells. Representative images of contracted gels (E) and quantification of the gel area after 4 days of contraction (F) are shown. Scale bar, 50 mm. Means ± SE (n=4); **P< 0.005; ***P< 0.0005, as compared to the control sgRNA group. A-F data are representative of three independent experiments.

Article Snippet: The following monoclonal (mAb) and polyclonal (pAb) antibodies were used to detect apical junction, cytoskeletal and matrix adhesion proteins: E-cadherin (610181), β-catenin (610154), mAbs (BD Biosciences, San Jose, CA); anti-cleaved PARP (9541), cleaved caspase3 (9664), NM-IIC (3405) pAbs (Cell Signaling, Beverly, MA); anti-ZO-1 (402200), phosphorylated paxillin (44722G) pAbs (Thermo Fisher Scientific, Waltham, MA); anti-occludin (13409), α-catenin (12831), GFP (50430) and anti-UNC-45A (19564) pAbs (Proteintech, Rosemont, IL); anti-E-cadherin (AF748) pAb (R & D System, Minneapolis, MN); NM IIA (909801) pAb (Biolegend, San Diego, CA); α-tubulin (T6199) mAb (Millipore-Sigma, San Louis, MO), anti-NM-IIA (H00004627-M03) mAb (Abnova, Walnut, CA).

Techniques: Microscopy, Immunofluorescence, Labeling

Journal: Cell Death Discovery

Article Title: Upregulation of CPNE7 in mesenchymal stromal cells promotes oral squamous cell carcinoma metastasis through the NF-κB pathway

doi: 10.1038/s41420-021-00684-w

Figure Lengend Snippet: Transwell assays were performed to detect tumour (CAL27 and WSU-HN6) cell migration and invasion through indirect cell-cell contact co-culture ( A ) and conditioned medium ( B ) from OLK-MSCs and OSCC-MSCs ( n = 5). C OLK-MSCs or OSCC-MSCs (10 6 ) were co-injected WSU-HN6 (10 6 ) into BALB/c mice via tail vein ( n = 6). Metastasis lung tumor nodules were counted after 28 days. OSCC-MSCs promote the metastasis of CAL27 ( C ) and WSU-HN6 cells ( D ) through epithelial‐mesenchymal transition (EMT)-related proteins (E‐cadherin, N‐cadherin, alpha E catenin, and Vimentin). The tests were repeated three times. Data were expressed as means ± SD. * p < 0.05.

Article Snippet: Equal amounts of proteins were resolved by 10% SDS-PAGE [ ] and immunoblotted with primary antibodies against the following antigens overnight: E-cadherin, alpha E catenin, Vimentin, N-cadherin, GFP (Proteintech, Wuhan, China), p-p65, total-p65 (Abcam, Cambridge, England), p-Iκβα, total-Iκβα (CST, California, USA), CPNE7 (GeneTex, Texas, USA), and GAPDH (Affinity, Ohio, USA), incubated with the HRP-conjugated secondary antibodies (Affinity, Ohio, USA).

Techniques: Migration, Co-Culture Assay, Injection

Journal: Balkan Medical Journal

Article Title: miR-509-3p Suppresses Migration, Invasion, and Epithelial–Mesenchymal Transition in Melanoma Cells by Targeting Collagen Triple Helix Repeat Containing 1

doi: 10.5152/balkanmedj.2021.20049

Figure Lengend Snippet: miR-509-3p suppressed epithelial–mesenchymal transition in melanoma cells. miR-509-3p mimic was transfected into SK-MEL-28 and WM115 cells, and protein expression levels of α-catenin, E-cadherin, vimentin, and fibronectin were detected by Western blot after 48 h of transfection. ** P < .01.

Article Snippet: The membrane was blocked in 5% skim milk and incubated with anti-CTHRC1 (1:1000; Proteintech Group, Chicago, IL, USA), anti-α-catenin (1:2000), anti-E-cadherin (1:2000), anti-vimentin (1:2500), anti-fibronectin (1:2500), or anti-β-actin (1:3000) antibodies.

Techniques: Transfection, Expressing, Western Blot

Journal: Balkan Medical Journal

Article Title: miR-509-3p Suppresses Migration, Invasion, and Epithelial–Mesenchymal Transition in Melanoma Cells by Targeting Collagen Triple Helix Repeat Containing 1

doi: 10.5152/balkanmedj.2021.20049

Figure Lengend Snippet: miR-509-3p suppressed malignant metastasis of melanoma cells through regulation of CTHRC1. (A) Effect of pcDNA-CTHRC1 and miR-509-3p mimic on protein expression of CTHRC1, α-catenin, E-cadherin, vimentin, and fibronectin in SK-MEL-28 and WM115 cells were detected by Western blot after 48 h of transfection. (B) Effect of pcDNA-CTHRC1 and miR-509-3p mimic on cell migration in SK-MEL-28 and WM115 cells were detected by wound healing after 48 h of transfection. (C) Effect of pcDNA-CTHRC1 and miR-509-3p mimic on cell invasion in SK-MEL-28 and WM115 cells were detected by transwell assay after 48 h of transfection. ** ,&& P < .01.

Article Snippet: The membrane was blocked in 5% skim milk and incubated with anti-CTHRC1 (1:1000; Proteintech Group, Chicago, IL, USA), anti-α-catenin (1:2000), anti-E-cadherin (1:2000), anti-vimentin (1:2500), anti-fibronectin (1:2500), or anti-β-actin (1:3000) antibodies.

Techniques: Expressing, Western Blot, Transfection, Migration, Transwell Assay