Chromatin Immunoprecipitation Assay: Examining the Interaction of NFkB with the VEGF Promoter

2015 ◽  
pp. 75-87 ◽  
Author(s):  
Chad B. Walton ◽  
Michelle L. Matter
Keyword(s):  
Chromatin Immunoprecipitation ◽  
Chromatin Immunoprecipitation Assay ◽  
Immunoprecipitation Assay

scholarly journals Transcriptional activation of CBFβ by CDK11p110 is necessary to promote osteosarcoma cell proliferation

2019 ◽  
Vol 17 (1) ◽  
Author(s):  
Yong Feng ◽  
Yunfei Liao ◽  
Jianming Zhang ◽  
Jacson Shen ◽  
Zengwu Shao ◽  
...  
Keyword(s):  
Chromatin Immunoprecipitation ◽  
Transcriptional Analysis ◽  
Luciferase Assay ◽  
Osteosarcoma Cell ◽  
Osteosarcoma Cells ◽  
Chromatin Immunoprecipitation Assay ◽  
Immunoprecipitation Assay ◽  
Gel Shift Assay ◽  
Gel Shift

Abstract Background Aberrant expression of cyclin-dependent protein kinases (CDK) is a hallmark of cancer. CDK11 plays a crucial role in cancer cell growth and proliferation. However, the molecular mechanisms of CDK11 and CDK11 transcriptionally regulated genes are largely unknown. Methods In this study, we performed a global transcriptional analysis using gene array technology to investigate the transcriptional role of CDK11 in osteosarcoma. The promoter luciferase assay, chromatin immunoprecipitation assay, and Gel Shift assay were used to identify direct transcriptional targets of CDK11. Clinical relevance and function of core-binding factor subunit beta (CBFβ) were further accessed in osteosarcoma. Results We identified a transcriptional role of protein-DNA interaction for CDK11p110, but not CDK11p58, in the regulation of CBFβ expression in osteosarcoma cells. The CBFβ promoter luciferase assay, chromatin immunoprecipitation assay, and Gel Shift assay confirmed that CBFβ is a direct transcriptional target of CDK11. High expression of CBFβ is associated with poor outcome in osteosarcoma patients. Expression of CBFβ contributes to the proliferation and metastatic behavior of osteosarcoma cells. Conclusions These data establish CBFβ as a mediator of CDK11p110 dependent oncogenesis and suggest that targeting the CDK11- CBFβ pathway may be a promising therapeutic strategy for osteosarcoma treatment. Graphical Abstract

A rapid micro chromatin immunoprecipitation assay (ChIP)

2008 ◽  
Vol 3 (6) ◽  
pp. 1032-1045 ◽  
Author(s):  
John Arne Dahl ◽  
Philippe Collas
Keyword(s):  
Chromatin Immunoprecipitation ◽  
Chromatin Immunoprecipitation Assay ◽  
Immunoprecipitation Assay

Studying KLF6 and PEPD ‐gene Promoter by Chromatin Immunoprecipitation Assay

2020 ◽  
Vol 34 (S1) ◽  
pp. 1-1
Author(s):  
Zeljka Miletic Lanaghan ◽  
Ireti Eni-Aganga ◽  
Muthukumar Balasubramaniam ◽  
Chandravanu Dash ◽  
Jui Pandhare
Keyword(s):  
Chromatin Immunoprecipitation ◽  
Gene Promoter ◽  
Chromatin Immunoprecipitation Assay ◽  
Immunoprecipitation Assay

scholarly journals MKP-1 mediates glucocorticoid-induced ERK1/2 dephosphorylation and reduction in pancreatic β-cell proliferation in islets from early lactating mothers

2010 ◽  
Vol 299 (6) ◽  
pp. E1006-E1015 ◽  
Author(s):  
José E. Nicoletti-Carvalho ◽  
Camilo Lellis-Santos ◽  
Tatiana S. Yamanaka ◽  
Tatiane C. Nogueira ◽  
Luciana C. Caperuto ◽  
...  
Keyword(s):  
Insulin Resistance ◽  
Pancreatic Islets ◽  
Chromatin Immunoprecipitation ◽  
Protein Phosphatase ◽  
Pancreatic Β Cell ◽  
Β Cell ◽  
Chromatin Immunoprecipitation Assay ◽  
Isolated Pancreatic Islets ◽  
Immunoprecipitation Assay ◽  
Lactating Mothers

Maternal pancreatic islets undergo a robust increase of mass and proliferation during pregnancy, which allows a compensation of gestational insulin resistance. Studies have described that this adaptation switches to a low proliferative status after the delivery. The mechanisms underlying this reversal are unknown, but the action of glucocorticoids (GCs) is believed to play an important role because GCs counteract the pregnancy-like effects of PRL on isolated pancreatic islets maintained in cell culture. Here, we demonstrate that ERK1/2 phosphorylation (phospho-ERK1/2) is increased in maternal rat islets isolated on the 19th day of pregnancy. Phospho-ERK1/2 status on the 3rd day after delivery (L3) rapidly turns to values lower than that found in virgin control rats (CTL). MKP-1, a protein phosphatase able to dephosphorylate ERK1/2, is increased in islets from L3 rats. Chromatin immunoprecipitation assay revealed that binding of glucocorticoid receptor (GR) to MKP-1 promoter is also increased in islets from L3 rats. In addition, dexamethasone (DEX) reduced phospho-ERK1/2 and increased MKP-1 expression in RINm5F and MIN-6 cells. Inhibition of transduction with cycloheximide and inhibition of phosphatases with orthovanadate efficiently blocked DEX-induced downregulation of phospho-ERK1/2. In addition, specific knockdown of MKP-1 with siRNA suppressed the downregulation of phospho-ERK1/2 and the reduction of proliferation induced by DEX. Altogether, our results indicate that downregulation of phospho-ERK1/2 is associated with reduction in proliferation found in islets of early lactating mothers. This mechanism is probably mediated by GC-induced MKP-1 expression.

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