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Journal: Cell Death & Disease
Article Title: CPNE7 promotes colorectal tumorigenesis by interacting with NONO to initiate ZFP42 transcription
doi: 10.1038/s41419-024-07288-z
Figure Lengend Snippet: A Volcano plots and Venn plot of DEGs in GEO CRC datasets GSE142279, GSE166254 and GSE196006. The selection criteria for DEGs were p -value < 0.05 and |FoldChange | >2. B GO analysis of common 1,354 DEGs in ( A ). BP: Biological Process, CC: Cellular Component, MF: Molecular Function. C KEGG analysis of common 1,354 DEGs in ( A ). D Heat map of the top 30 genes from 647 common upregulated DEGs in ( A ). E , F CPNE7 expression in COAD, READ and normal tissues in UALCAN database ( E ) and GEPIA database ( F ). TPM: Transcripts Per Million. G CPNE7 expression in 19 pairs of CRC samples were detected by qRT-PCR. H , I CPNE7 mRNA expression in normal tissues, tumor tissues and liver metastatic tissues measured by RNA-sequencing ( H ) and qRT-PCR ( I ) in paired CRC samples. J Immunohistochemical staining of CPNE7 in normal tissues, tumor tissues and liver metastatic tissues of CRC samples. Scale bar: 100 μm. K Relationship between CPNE7 expression and CRC patients’ survival in HPA database. For ( G ), two-tailed paired Student’s t -test was used. For ( I ), data are shown as mean ± SD and two-tailed unpaired Student’s t -test was used. * p < 0.05, ** p < 0.01, *** p < 0.001. Data are representative of at least three independent experiments.
Article Snippet: The following antibodies were used in our study: Anti-CPNE7 antibody (Cat# bs-14030R, Bioss),
Techniques: Selection, Expressing, Quantitative RT-PCR, RNA Sequencing, Immunohistochemical staining, Staining, Two Tailed Test
Journal: Cell Death & Disease
Article Title: CPNE7 promotes colorectal tumorigenesis by interacting with NONO to initiate ZFP42 transcription
doi: 10.1038/s41419-024-07288-z
Figure Lengend Snippet: A , B Expression of mRNA ( A ) and protein ( B ) of CPNE7 in human normal colonic epithelial cell line NCM460 and human CRC tumor cell lines HCT116, SW620, SW480 and HT29. C , D Expression of CPNE7 at the mRNA level ( C ) and protein level ( D ) in HCT116 and SW620 cell lines. E Cell viability was measured by CCK-8 assay. F Colony formation assays of shControl and shCPNE7 CRC cells. Representative images are shown on the left, and the statistical analysis are is on the right. G Transwell assays of shControl and shCPNE7 CRC cells. Representative images are shown on the left, and the statistical analysis is shown on the right. H Apoptosis detection assays of shControl and shCPNE7 CRC cells. Representative images are shown on the left, and the statistical analysis of apoptotic rates (including early apoptosis and late apoptosis) is shown on the right. For ( A – H ), data are shown as mean ± SD and two-tailed unpaired Student’s t -test was used. * p < 0.05, ** p < 0.01, *** p < 0.001. Data are re p resentative of at least three independent experiments.
Article Snippet: The following antibodies were used in our study: Anti-CPNE7 antibody (Cat# bs-14030R, Bioss),
Techniques: Expressing, CCK-8 Assay, Two Tailed Test
Journal: Cell Death & Disease
Article Title: CPNE7 promotes colorectal tumorigenesis by interacting with NONO to initiate ZFP42 transcription
doi: 10.1038/s41419-024-07288-z
Figure Lengend Snippet: A shControl and shCPNE7 HCT116 cells were injected into BALB/c-nude mice ( n = 5 mice per group). Volume changes of xenograft tumors in nude mice are shown on the left, and mice images are shown on the right. B Tumor images are shown on the left, and statistical analysis for tumor weights is shown on the right. C Expression of CPNE7 in xenograft tumors were detected by Western blot. D IHC staining of Ki-67 in xenograft tumors in ( B ). Scale bar: 100 μm. E Representative images of the visible metastatic nodules in livers. ( n = 4 mice per group). F Representative images of liver ( E ) histology stained with hematoxylin and eosin are shown on the left, and the statistical analysis is shown on the right. For ( A – F ), data are shown as mean ± SD and two-tailed unpaired Student’s t -test was used. * p < 0.05, ** p < 0.01, *** p < 0.001. Data are re p resentative of at least three independent experiments.
Article Snippet: The following antibodies were used in our study: Anti-CPNE7 antibody (Cat# bs-14030R, Bioss),
Techniques: Injection, Expressing, Western Blot, Immunohistochemistry, Staining, Two Tailed Test
Journal: Cell Death & Disease
Article Title: CPNE7 promotes colorectal tumorigenesis by interacting with NONO to initiate ZFP42 transcription
doi: 10.1038/s41419-024-07288-z
Figure Lengend Snippet: A CPNE7 overexpression HT29 cell line was established and verified by Western blot. B Cell viability was measured by CCK-8 assay. C Colony formation assays of Vector and oeCPNE7 HCT116 cells. Representative images are shown on the left, and the statistical analysis is shown on the right. D Transwell assays of Vector and oeCPNE7 HCT116 cells. Representative images are shown on the left, and the statistical analysis is shown on the right. E Wound-healing assays of Vector and oeCPNE7 HCT116 cells. Representative images are shown on the left, and the statistical analysis is shown on the right. F Vector and oeCPNE7 HT29 cells were injected into BALB/c-nude mice ( n = 5 mice per group). Volume changes of xenograft tumors in nude mice are shown. G Tumor images are shown on the left, and statistical analysis for tumor weights is shown on the right. H Expression of CPNE7 in xenograft tumors were detected by qRT-PCR. I IHC staining of CPNE7 and Ki-67 in xenograft tumors in ( G ). Scale bar: 100 μm. For ( B – G ), data are shown as mean ± SD and two-tailed unpaired Student’s t -test was used. * p < 0.05, ** p < 0.01, *** p < 0.001. Data are re p resentative of at least three independent experiments.
Article Snippet: The following antibodies were used in our study: Anti-CPNE7 antibody (Cat# bs-14030R, Bioss),
Techniques: Over Expression, Western Blot, CCK-8 Assay, Plasmid Preparation, Injection, Expressing, Quantitative RT-PCR, Immunohistochemistry, Two Tailed Test
Journal: Cell Death & Disease
Article Title: CPNE7 promotes colorectal tumorigenesis by interacting with NONO to initiate ZFP42 transcription
doi: 10.1038/s41419-024-07288-z
Figure Lengend Snippet: A Volcano plots of DEGs in shControl group vs shCPNE7 groups. B , C GO and KEGG analysis of DEGs from left volcano plot ( B ) and right volcano plot ( C ) in ( A ). D GSEA analysis of RNA-sequencing data from shControl group and shCPNE7-1 group. E Venn plot of HumanTFDB and DEGs in RNA-sequencing (|FoldChange | > 1.5). F 5 genes exhibiting the same trend in RNA-sequencing and qRT-PCR. G Immunofluorescence staining of CPNE7 and NONO in HCT116 cells. Scale bar: 10 μm. H Dual-luciferase reporter assay were performed in shControl and shCPNE7 293 T cells. I Proteins from the IP assay on SW480 cells were separated by SDS-PAGE and detected by silver staining. J , K Interaction between NONO and CPNE7 were verified in 293 T ( J ) and HCT116 ( K ) cells by Co-IP assay. L The effect of CPNE7 and NONO on ZFP42 promoter activity was detected by dual-luciferase reporter assay. M Left panel: schematic diagram of the structure of truncated ZFP42 promoter reporter plasmids. Right panel: Changes in luciferase activity following truncation at different positions of the ZFP42 promoter. N ZFP42 mRNA expression were detected in shControl and shNONO SW620 cells. O Expression of NONO and ZFP42 were detected in CPNE7 knockout HCT116 cells by Western blot. P , Q mRNA expression level ( P ) and protein expression level ( Q ) of ZFP42 in shControl and shZFP42 HCT116 cells. R Cell viability was measured by CCK-8 assay. S Colony formation assays of shControl and shZFP42 HCT116 cells. Representative images are shown on the left, and the statistical analysis is shown on the right. T Transwell assays of shControl and shZFP42 HCT116 cells. Representative images are shown on the left, and the statistical analysis is shown on the right. U Wound-healing assays of shControl and shZFP42 HCT116 cells. Representative images are shown on the left, and the statistical analysis for migration rates is shown on the right. V Apoptosis detection assays of shControl and shZFP42 HCT116 cells. Representative images are shown on the left, and the statistical analysis for apoptotic rates (including early apoptosis and late apoptosis) is shown on the right. For ( F – V ), data are shown as mean ± SD and two-tailed unpaired Student’s t -test was used. * p < 0.05, ** p < 0.01, *** p < 0.001. Data are representative of at least three independent experiments.
Article Snippet: The following antibodies were used in our study: Anti-CPNE7 antibody (Cat# bs-14030R, Bioss),
Techniques: RNA Sequencing, Quantitative RT-PCR, Immunofluorescence, Staining, Luciferase, Reporter Assay, SDS Page, Silver Staining, Co-Immunoprecipitation Assay, Activity Assay, Expressing, Knock-Out, Western Blot, CCK-8 Assay, Migration, Two Tailed Test
Journal: Cell Death & Disease
Article Title: CPNE7 promotes colorectal tumorigenesis by interacting with NONO to initiate ZFP42 transcription
doi: 10.1038/s41419-024-07288-z
Figure Lengend Snippet: A Schematic of CPNE7 shRNA intratumoral injection. B , C Tumor volumes ( B ) and tumor weights ( C ) of two groups. ( n = 4 tumors per group). D IHC staining of CPNE7 and Ki-67 in tumors in ( A ). Scale bar: 100 μm. E Protein expression level of CPNE7 in ( A ) was detected by Western blot. F Schematic of protein docking between CPNE7 and NONO. G Top 10 compounds with the lowest S score obtained from virtual screening. H Compound toxicity was tested by CCK-8 assay. I IC50 determination of Gramicidin in HCT116 and SW620 cells. J Disruption of Gramicidin on CPNE7-NONO interaction was verified by Co-IP assay. The concentration of Gramicidin was 8 nM. K The expression of ZFP42 mRNA in SW620 was detected after treatment with Gramicidin. L Schematic of Gramicidin intratumoral injection. M Volume changes of subcutaneous tumors in nude mice are shown on the left, and mice image are shown on the right ( n = 5 tumors per group). N Tumor images are shown on the left, and statistical analysis for tumor weights is shown on the right. O Tumor inhibition rate. P The expression of ZFP42 mRNA in tumors was detected by qRT-PCR. Q IHC staining of Ki-67 in tumors in ( M ). Scale bar: 100 μm. For ( B – Q ), data are shown as mean ± SD and two-tailed unpaired Student’s t -test was used. * p < 0.05, ** p < 0.01, *** p < 0.001. For ( H – Q ), data are representative of at least three independent experiments.
Article Snippet: The following antibodies were used in our study: Anti-CPNE7 antibody (Cat# bs-14030R, Bioss),
Techniques: shRNA, Injection, Immunohistochemistry, Expressing, Western Blot, CCK-8 Assay, Disruption, Co-Immunoprecipitation Assay, Concentration Assay, Inhibition, Quantitative RT-PCR, Two Tailed Test