Journal: bioRxiv

Article Title: KLF7-Regulated ITGA2 as a Therapeutic Target for Inhibiting Oral Cancer Stem Cells

doi: 10.1101/2024.11.04.621805

Figure Lengend Snippet: A The log2-transformed fold change (FC) in RNA levels between the stem-like cluster with the rest of the clusters and the stem-related pathway with the rest of the pathway. B Immunoblot assay of ITGA2, phosphor-ERK1/2, total-ERK1/2, phosphor-AKT, total-AKT, phosphor-YAP1, total-YAP1 protein levels in CAL27 and HSC3 cells after stable silencing ITGA2. C Representative immunofluorescent image showing the expression of YAP1 and DAPI in wt and stable silencing ITGA2 cells. D The sequence and domain in ITGA2 protein. E Co-IP analysis in CAL27 and HSC3 cells transduced with ITGA2 OE and ITGA2 mut plasmid. F Immunoblot assay of phosphor-ERK1/2, total-ERK1/2, phosphor-AKT, total-AKT, phosphor-YAP1, total-YAP1 protein levels in CAL27 and HSC3 cells after transduced with ITGA2 OE and ITGA2 mut plasmid. G Representative immunofluorescent image showing the expression of YAP1 and DAPI in CAL27 and HSC3 cells after transduced with ITGA2 OE and ITGA2 mut plasmid.

Article Snippet: After washing and blocking, the primary antibody against YAP1 (66900-1-Ig, Proteintech) was incubated overnight.

Techniques: Transformation Assay, Western Blot, Expressing, Sequencing, Co-Immunoprecipitation Assay, Transduction, Plasmid Preparation

Journal: bioRxiv

Article Title: KLF7-Regulated ITGA2 as a Therapeutic Target for Inhibiting Oral Cancer Stem Cells

doi: 10.1101/2024.11.04.621805

Figure Lengend Snippet: A The molecular formula of TC-I 15. B Co-IP analysis in CAL27 and HSC3 cells treated with DMSO and TC-I 15. C CAL27 cells were intracardially injected into mice. Seven days later, TC-I 15 was injected (20 mg/kg) into mice via vein every third day for 1 month. D Representative images showing the xenograft model in CA27 cells treated with DMSO or TC-I 15(n = 5 per group) and the growth of tumor grafts was shown. E Immunoblot assay of phosphor-ERK1/2, total-ERK1/2, phosphor-AKT, total-AKT, phosphor-YAP1, and total-YAP1 protein levels in xenograft tissue. F Representative FACS plots and quantification of CD133+cells in the xenograft tissues. G CAL27 cells were intracardially injected into mice. Seven days later, TC-I 15 (20 mg/kg) and plastin(5 mg/kg) were injected into nude mice via vein every third day for 1 month. H Representative images showing the xenograft model in CA27 cells treated with DMSO or TC-I 15(n = 5 per group) and the growth of tumor grafts were shown.

Article Snippet: After washing and blocking, the primary antibody against YAP1 (66900-1-Ig, Proteintech) was incubated overnight.

Techniques: Co-Immunoprecipitation Assay, Injection, Western Blot

Journal: Bone Research

Article Title: Trim21 depletion alleviates bone loss in osteoporosis via activation of YAP1/β-catenin signaling

doi: 10.1038/s41413-023-00296-3

Figure Lengend Snippet: YAP1/β-catenin signaling is essential for Trim21-mediated osteogenic differentiation. a Schematic diagram showing TMT-based quantitative proteomics for the identification of differentially expressed proteins (DEPs) in bone marrow mesenchymal stem cells (BMSCs) derived from Trim21 +/+ and Trim21 −/− mice. b Volcano plots of DEPs in BMSCs from Trim21 +/+ and Trim21 −/− mice. c Heatmap analysis of DEPs in BMSCs. Three replicates of each group were included, and the top 29 DEPs are shown. d KEGG enrichment analysis of the DEPs in the BMSCs. e Representative immunoblotting analysis and quantification of BCL9, AXIN1, and YAP1 in BMSCs; proteomics sample: part of the samples subjected to proteomics analysis. f Representative immunoblotting analysis and quantification of BCL9, β-catenin, YAP1, and Runx2 protein expression in BMSCs after osteogenic induction for 7 days. g The endogenous interaction between Trim21, BCL9, β-catenin, and YAP1 was evaluated using a co-IP assay. h Protein‒protein interaction of YAP1 and Trim21 in living cells. The two BiFC plasmids encoding Myc-VN155-YAP1 and HA-VC155-Trim21 along with HA-cerulean were cotransfected into HEK293T cells for 24 h. Representative images showing transfected cells (cerulean) and the interaction between YAP1 and Trim21 (Venus). Nuclei were stained with DAPI. Scale bar: 20 μm. i Immunoblotting analysis of BCL9, β-catenin, YAP1, and HA-Trim21 protein expression in HEK293T cells treated with or without MG132. j Representative images showing the expression of YAP1 + OBs derived from Trim21 +/+ and Trim21 −/ − mice. Scale bar: 20 μm. All bar graphs are presented as the mean ± SD. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.000 1; n.s. not significant by Student’s t test

Article Snippet: Next, 1% goat serum was used to block tissues at 37 °C for 30 min. For IHC staining, sections were treated with primary antibody against YAP1 (CST, Cat# 14074, USA, 1:200) overnight at 4 °C.

Techniques: Quantitative Proteomics, Derivative Assay, Western Blot, Expressing, Co-Immunoprecipitation Assay, Transfection, Staining

Journal: Bone Research

Article Title: Trim21 depletion alleviates bone loss in osteoporosis via activation of YAP1/β-catenin signaling

doi: 10.1038/s41413-023-00296-3

Figure Lengend Snippet: Trim21 orchestrates ovariectomy (OVX)-induced bone metabolism by targeting YAP1 signaling. a Determination of fat cell density in the proximal tibia of 20-week-old Trim21 +/+ and Trim21 −/− mice induced by sham operation or OVX. b Representative micro-CT images and quantitative data (BV/TV) of proximal tibial bone of 20-week-old Trim21 +/+ and Trim21 −/− mice induced by sham operation or OVX. c , d Calcein double labeling of mineral layers of tibial trabecular bone of 5-month-old mice. e Representative images of von Kossa staining of the undecalcified proximal tibia of 5-month-old mice. Scale bar: 50 μm. f IHC staining images of the proximal tibia of 5-month-old mice using an antibody against YAP1. The YAP1-stained positive cells are denoted by the red arrow. Scale bar: 50 μm. g , h Representative images of histological sections of the tibia that were stained with TRAP in Trim21 +/+ and Trim21 −/− mice induced by sham operation or OVX. TRAP-stained osteoclasts (OCs) are denoted by the red arrow. OC. N/BPm (OC number per bone parameter) and OC. S/BS (OC surface per bone surface) was determined. Scale bar: 100 μm. All bar graphs are presented as the mean ± SD. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.000 1; n.s. not significant by Student’s t test

Article Snippet: Next, 1% goat serum was used to block tissues at 37 °C for 30 min. For IHC staining, sections were treated with primary antibody against YAP1 (CST, Cat# 14074, USA, 1:200) overnight at 4 °C.

Techniques: Micro-CT, Labeling, Staining, Immunohistochemistry

Journal: Bone Research

Article Title: Trim21 depletion alleviates bone loss in osteoporosis via activation of YAP1/β-catenin signaling

doi: 10.1038/s41413-023-00296-3

Figure Lengend Snippet: A schematic of Trim21 in the regulation of bone remodeling via YAP1/β-catenin signaling. Normal bone remodeling is maintained by the balance of MSC/osteoblast-mediated bone formation and osteoclast-mediated bone resorption. Trim21, by interacting with the protein complex formed by YAP1/β-catenin/BCL9, dictates the degradation of this protein complex, which in turn inactivates YAP1 and β-catenin signaling, which is essential for osteoblast differentiation. However, Trim21 is critical for maintaining the basic expression of osteoclast biomarkers, including Nfatc1 and Ctsk . Therefore, the coupling of osteoblasts with osteoclasts is attributed to dynamic changes in bone metabolism (left panel). In contrast, the loss of Trim21 causes disassociation with the YAP1/β-catenin/BCL9 complex, which then enters the nucleus for subsequent activation of osteogenic genes, including Runx2 and Osterix . In addition, the loss of Trim21 suppresses the maturation of osteoclasts. Together, these results indicate that Trim21 deficiency alleviates pathological bone loss by activating YAP1/β-catenin signaling

Article Snippet: Next, 1% goat serum was used to block tissues at 37 °C for 30 min. For IHC staining, sections were treated with primary antibody against YAP1 (CST, Cat# 14074, USA, 1:200) overnight at 4 °C.

Techniques: Expressing, Activation Assay

Journal: Cancer Biology & Therapy

Article Title: miR-506 is a YAP1-dependent tumor suppressor in laryngeal squamous cell carcinoma

doi: 10.1080/15384047.2018.1564569

Figure Lengend Snippet: Categorization of LSCC patients and statistical analysis of their correlations with the expression levels of miR-506 and YAP1.

Article Snippet: The primary antibodies against YAP1 and caspase-3 were obtained from Cell Signaling Technology, Inc..

Techniques: Expressing

Journal: Cancer Biology & Therapy

Article Title: miR-506 is a YAP1-dependent tumor suppressor in laryngeal squamous cell carcinoma

doi: 10.1080/15384047.2018.1564569

Figure Lengend Snippet: Correlations between miR-506 and Yap1 in LSCC patients.

Article Snippet: The primary antibodies against YAP1 and caspase-3 were obtained from Cell Signaling Technology, Inc..

Techniques: Expressing

Journal: Scientific Reports

Article Title: Establishment of well-differentiated camelid airway cultures to study Middle East respiratory syndrome coronavirus

doi: 10.1038/s41598-022-13777-y

Figure Lengend Snippet: Establishment and characterization of Bactrian camel and llama AEC cultures. ( A ) Immunofluorescence analysis showing the development of tight-junctions (ZO-1, white) and ciliogenesis (β-tubulin, red) in Bactrian camel and llama AEC cultures over time from 1-day to 4 weeks post ALI exposure. The cells were counterstained with DAPI (blue) to visualize the nuclei. ( B,C ) Ciliogenesis quantification of camel and llama AEC cultures overtime, respectively. Ciliation was quantified by measuring the area above a fluorescence intensity threshold of five random images acquired per condition. ( D ) Transepithelial electrical resistance (TEER) measurement of camel and llama AEC cultures overtime during the differentiation. ( E ) Epithelial morphology of ex vivo tissues (upper panel) and well-differentiated camel and llama AEC cultures (lower panel). ( F ) DPP4 expression in well-differentiated camel and llama AEC cultures, with Vero cells as a positive control. Scale bar is 20 µm.

Article Snippet: Alexa-Fluor ® 647-labelled rabbit anti β-tubulin IV (Cell Signalling Technology, 9F3) and Alexa-Fluor ® 594-labelled mouse antibody against ZO-1 (Thermo Fisher Scientific, 1A12) were used to visualize cilia and tight junctions, respectively.

Techniques: Immunofluorescence, Fluorescence, Ex Vivo, Expressing, Positive Control

Journal: Iranian Journal of Pathology

Article Title: YAP1 and P53 Expression in Papillary Thyroid Carcinoma

doi: 10.30699/IJP.2023.553716.2897

Figure Lengend Snippet: Immunohistochemical expression of YAP1 in PTC; (A) positive cytoplasmic staining X10, (B)negative staining X40

Article Snippet: The slices were incubated with primary antibodies against YAP1 (1:200, clone (G-6), Santa Cruz Biotechnology, Dallas, Texas, USA) and p53 (1:1000, clone DO-7; Santa Cruz Biotechnology, Dallas, Texas, USA).

Techniques: Immunohistochemical staining, Expressing, Staining, Negative Staining

Journal: Iranian Journal of Pathology

Article Title: YAP1 and P53 Expression in Papillary Thyroid Carcinoma

doi: 10.30699/IJP.2023.553716.2897

Figure Lengend Snippet: PTC patients' Characteristics According to the immunohistochemical expression of YAP1

Article Snippet: The slices were incubated with primary antibodies against YAP1 (1:200, clone (G-6), Santa Cruz Biotechnology, Dallas, Texas, USA) and p53 (1:1000, clone DO-7; Santa Cruz Biotechnology, Dallas, Texas, USA).

Techniques: Immunohistochemical staining, Expressing