anti cysteine rich angiogenic inducer 61 (Danaher Inc)
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Anti Cysteine Rich Angiogenic Inducer 61, supplied by Danaher Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "YAP1-CPNE3 positive feedback pathway promotes gastric cancer cell progression"
Article Title: YAP1-CPNE3 positive feedback pathway promotes gastric cancer cell progression
Journal: Cellular and Molecular Life Sciences
doi: 10.1007/s00018-024-05178-3
Figure Legend Snippet: Screening of CPNE3 as a downstream target gene regulated by YAP1. A mRNA sequencing in BGC-823 cells with downregulated YAP1 expression. B Examination of mRNA expression by quantitative reverse transcription polymerase chain reaction (RT-qPCR) after downregulating YAP1 in BGC-823 cells. C , D mRNA expression levels of YAP1 , CPNE3 , and CYR61 in AGS and HGC-27 cells (transfected with NC, siYAP1-#1, and siYAP1-#2 siRNAs). E Two-dimensional visualization of CPNE3 and YAP1 in single-cell clusters in patients with gastric cancer (GC). F Protein expression levels of YAP1, CPNE3, and CYR61 following down-regulation of YAP1 expression in AGS and HGC-27 cells. G , H After gradient overexpression of Flag-YAP1, the YAP1, CPNE3, and CYR61 proteins and mRNA levels were detected by western blotting (WB) and RT-qPCR, respectively. Three independent biological experiments were performed, and statistical significance is denoted by * p < 0.05 and ** p < 0.01
Techniques Used: Sequencing, Expressing, Reverse Transcription Polymerase Chain Reaction, Quantitative RT-PCR, Transfection, Over Expression, Western Blot
Figure Legend Snippet: CPNE3 is associated with the Hippo-YAP1 pathway in GC. A Enrichment analysis of CPNE3 with Hippo-YAP1. B Pearson correlation analyses of CPNE3 with Hippo-YAP1 C – E Using siRNA, the expression of CPNE3 at the mRNA and protein levels were downregulated in BGC-823 and MKN-28 cells. F , G Up-regulation of CPNE3 expression by transfection of HA-CPNE3 plasmid in AGS and HGC-27 cells. H WB revealed that the expression of YAP1 and its target gene CYR61 were both considerably upregulated in AGS and HGC-27 cells using the same technique used to boost CPNE3 expression as above. I , J Confocal images of HA-CPNE3 plasmid transfection in HGC-27 cells and siCPNE3-#1 or siCPNE3-#2 transfection in BGC-823 cells with YAP1 labeling
Techniques Used: Expressing, Transfection, Plasmid Preparation, Labeling
Figure Legend Snippet: CPNE3 promotes GC progression in a partial YAP1-dependent manner. BGC-823 and MKN-28 cells were treated with the ShCPNE3-#2 plasmid to downregulate the expression of CPNE3 and the Flag-YAP1 plasmid to concurrently increase the expression of YAP1. A WB was used to measure the expression of CPNE3, YAP1, and CYR61. B – G Phenotyping assays were used to determine the degree to which the CPNE3 depletion-induced suppression of GC cell proliferation, migration, invasion, colony formation, and drug resistance could be reversed by the overexpression of exogenous YAP1. H – K The capacity of MKN-45 cells to proliferate, invade, migrate, and form colonies was weakly inhibited by downregulation of CPNE3 expression in MKN-45 cells lacking YAP1 expression. ShYAP1-#1 plasmid-mediated stable YAP1 knockdown or HA-CPNE3 plasmid-mediated simultaneous overexpression of CPNE3 in AGS and HGC-27 cells. L WB was used to investigate the relevant protein levels. M – P The overexpression of CPNE3 did not alleviate the suppression of GC cell proliferation, migration, invasion, and colony formation brought on by the downregulation of YAP1. Three independent biological experiments were conducted, and statistical significance is shown by the notations, * p < 0.05 and ** p < 0.01
Techniques Used: Plasmid Preparation, Expressing, Migration, Over Expression
Figure Legend Snippet: CPNE3 promotes GC growth in vivo. A , B Cell line with stable silencing of CPNE3 expression was established in BGC-823 cells by infection with lentivirus encoding sgRNA targeting CPNE3 or negative control. C – E Silencing of CPNE3 significantly reduced tumor growth in vivo, and the weight and volume of the tumor tissues were significantly lower than those in controls (mean ± standard error of the mean [SEM] n = 10/group). F , G Immunohistochemistry (IHC) and WB tests were used to assess the protein levels of CPNE3, YAP1, and CYR61 in tumor tissues from subcutaneous cell line-derived xenograft models constructed from BGC-823 cells with CPNE3 expression downregulated by lentivirus. H – J Using the same method described above to validate the function of CPNE3 in a patient-derived xenograft (PDX) model of GC, silencing CPNE3 significantly reduced tumor growth, weight, and volume in vivo (mean ± SEM n = 6/group). K , L Protein expression of CPNE3, YAP1, and CYR61 were detected using WB and IHC, assays in BGC-823 cells after stably downregulating CPNE3 expression in the PDX model
Techniques Used: In Vivo, Expressing, Infection, Negative Control, Immunohistochemistry, Derivative Assay, Construct, Stable Transfection
Figure Legend Snippet: CPNE3 is an independent prognostic factor that causes poor prognosis in patients with GC. A Using Western blotting, the protein levels of CPNE3 in eight pairs of GC patient tissue samples were measured. B CPNE3 protein expression was examined the IHC of tumor tissues (n = 20) and matched normal tissues (n = 20) from patients with GC. C – E Overall survival and CPNE3 expression were used to stratify the patients in the training, validation, and training + validation groups, followed by Kaplan–Meier analysis. F The chi-square test was used to determine the relationship between the expression of CPNE3 and that of YAP1, CYR61, and RAD51. G Kaplan–Meier survival analysis was performed in 100 patients who were categorized based on their CPNE3 and YAP1 protein levels. H Diagram by Figdraw showing the process through which the CPNE3-YAP1 positive feedback loop promotes GC
Techniques Used: Western Blot, Expressing