A Simplified Method for the Preparation of Transcriptionally Active Liver Nuclear Extracts

Hattori, Masahira; Tugores, Antonio; Veloz, Linda; Karin, Michael; Brenner, David A.

doi:10.1089/dna.1990.9.777

Cited by 126 publications

(66 citation statements)

References 23 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Additional double-stranded oligonucleotides used as competitors and/or probes included NFkB-CONS (5Ј-AGTTGAGGGGACTTTCCCAGGC-3Ј), which contains the NFB consensus binding site, AP2CONS (5Ј-GATCGAACTGACCGC-CCGCGGCCCGT-3Ј), which contains the human metallothionein IIa promoter AP-2-binding site (22), and AML1CONS (5Ј-GGATATTT-GCGGTTAGCA-3Ј) (23). DNase I protection was performed as described (24), except that cells were lysed in 10 mM Hepes, pH 7.6, 10 mM KCl, 0.1 mM EDTA, 0.1 mM EGTA, 0.75 mM spermidine, 0.1 mM spermine, 2 g/ml aprotinin, 2 g/ml leupeptin, and 10 mM Na 2 MoO 4 . Samples were subjected to electrophoresis on 6% denaturing polyacrylamide gels in parallel with a G ϩ A sequence ladder generated with the same probe.…”

Section: Methodsmentioning

confidence: 99%

Polyomavirus Enhancer-binding Protein 2/Core Binding Factor/Acute Myeloid Leukemia Factors Contribute to the Cell Type-specific Activity of the CD11a Integrin Gene Promoter

Puig‐Kröger¹,

López-Rodrı́guez²,

Relloso³

et al. 2000

Journal of Biological Chemistry

View full text Add to dashboard Cite

The CD11a/CD18 leukocyte integrin (LFA-1; also known as ␣L/␤2) mediates leukocyte transendothelial migration during immune and inflammatory responses and participates in lymphoma metastasis. CD11a/CD18 leukocyte-restricted expression is controlled by the CD11a gene promoter, which confers tissue-specific expression to reporter genes in vitro and in vivo. DNase I protection analysis of the CD11a proximal gene promoter revealed DNA-protein interactions centered at position -110 (CD11a-110). Disruption of CD11a-110 reduced CD11a promoter activity in a cell type-specific manner, as it reduced its activity by 70% in Jurkat lymphoid cells, whereas the effect was considerably lower in K562 and HepG2 cells. Electrophoretic mobility shift assays showed evidence of cell type-specific differences in CD11a-110 binding and indicated its specific recognition by members of the polyomavirus enhancerbinding protein 2/core binding factor (CBF)/acute myeloid leukemia (AML) family of transcription factors. AML1B/CBF␤ transactivated the CD11a promoter, with AML1B/CBF␤-mediated transactivation being completely dependent on the integrity of the CD11a-110 element. Therefore, CBF/AML factors play a role in the cell type-restricted transcription of the CD11a integrin gene through recognition of CD11a-110. The involvement of CBF/AML factors in CD11a expression raises the possibility that CD11a/CD18 expression might be deregulated in acute myeloid and B-lineage acute lymphoblastic leukemias, thus contributing to their altered adhesion and metastatic potential.

show abstract

Section: Methodsmentioning

confidence: 99%

Polyomavirus Enhancer-binding Protein 2/Core Binding Factor/Acute Myeloid Leukemia Factors Contribute to the Cell Type-specific Activity of the CD11a Integrin Gene Promoter

Puig‐Kröger¹,

López-Rodrı́guez²,

Relloso³

et al. 2000

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…Nuclear proteins were prepared from frozen livers using ultracentrifugation as described ( 24,25 ). Equal aliquots (20 g) of nuclear protein from individual livers were subjected to SDS-PAGE on 4-12% gels and transferred to a polyvinylidene difl uoride membrane.…”

Section: Western Blot Analysismentioning

confidence: 99%

Botanical oils enriched in n-6 and n-3 FADS2 products are equally effective in preventing atherosclerosis and fatty liver

Shewale

Boudyguina

Zhu

et al. 2015

Journal of Lipid Research

View full text Add to dashboard Cite

“…Nuclear proteins were prepared from liver tissues as previously described (12). Binding reactions were carried out for 20 min on ice using 8 g nuclear extract and 10 pg [ 32 P]DNA probe containing a NF-B consensus binding site (3) with or without 1 ng (100-fold) unlabeled competitor probe.…”

Section: Nuclear Extract Preparation and Mobility Shift Assaymentioning

confidence: 99%

NF-κB activation in Kupffer cells after partial hepatectomy

Yang

Magness

Bataller

et al. 2005

American Journal of Physiology-Gastrointestinal and Liver Physi

Self Cite

View full text Add to dashboard Cite

The transcription factor nuclear factor-κB (NF-κB) is activated during liver regeneration after partial hepatectomy. However, the physiological role and cellular localization of NF-κB activation are unresolved. In this study, we used an adenoviral vector expressing a mutated form of IκBα to inhibit NF-κB activity during liver regeneration. After partial hepatectomy in mice, introduction of Ad5IκB, but not a control virus (Ad5GFP), resulted in increased liver injury and decreased hepatocyte proliferation. Hepatocyte apoptosis was not observed. To investigate the kinetics and cellular localization of NF-κB-induced transcription during liver regeneration, we generated a transgenic mouse expressing enhanced green fluorescent protein (EGFP) under the transcriptional control of NF-κB cis elements ( cis-NF-κB-EGFP). During liver regeneration, EGFP expression was detected within 12 h and was primarily located in Kupffer cells. Our data demonstrate that activation of NF-κB initially occurs in Kupffer cells after partial hepatectomy in mice.

show abstract

A Simplified Method for the Preparation of Transcriptionally Active Liver Nuclear Extracts

Cited by 126 publications

References 23 publications

Polyomavirus Enhancer-binding Protein 2/Core Binding Factor/Acute Myeloid Leukemia Factors Contribute to the Cell Type-specific Activity of the CD11a Integrin Gene Promoter

Polyomavirus Enhancer-binding Protein 2/Core Binding Factor/Acute Myeloid Leukemia Factors Contribute to the Cell Type-specific Activity of the CD11a Integrin Gene Promoter

Botanical oils enriched in n-6 and n-3 FADS2 products are equally effective in preventing atherosclerosis and fatty liver

NF-κB activation in Kupffer cells after partial hepatectomy

Contact Info

Product

Resources

About