Influence of notch dependent signaling on NF-kappaB activity and apoptosis of myelocytic cells:
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| Main Author: | |
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| Format: | Thesis Book |
| Language: | English |
| Published: |
2004
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| Subjects: | |
| Online Access: | Inhaltsverzeichnis |
| Physical Description: | 96 S. Ill., graph. Darst. |
Staff View
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| 245 | 1 | 0 | |a Influence of notch dependent signaling on NF-kappaB activity and apoptosis of myelocytic cells |c von Aneta Dobierzewska |
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Record in the Search Index
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Table of Context
1. Zusammenfassung/Summary 3
2. Introduction 7
2.1. Notch Receptors and their Ligands 8
2.2. Structure of Notch Proteins Family 10
2.3. Principles of Notch Signaling 12
2.4. Target Genes in Notch Signaling Pathway 13
2.5. Regulation of Notch Signaling 15
2.6. Expression of Members of Notch Family in Hematopoietical Cells 15
2.7. NF-kB Proteins Family 17
2.8. NF-kB Transduction Pathway 19
2.9. 'Cross-talk' between Notch and NF-kB Signaling Pathways 22
2.10. Influence of Notch Signaling on Apoptotic Cell Death 23
2.11. Scope of the Thesis 25
3. Results 26
3.1.Notch-l andNotch-2 receptors are expressed in myeloid cells 26
3.2. The Notch-1 and Notch-2 retroviral constructs used for retrovirus-mediated gene
transfer in 32D cells 28
33. Retroviral Infection and Subcloning of 32D cells producing Notch- 1-IC and
Notch-2-IC 30
3.4. Influence of Notch-1-IC and Notch-2-IC on intracellular Notch signaling pathways. .33
3.5. LPS-induced NF-kB promoter activity is inhibited by Notch-1 and stimulated by
Notch-2 34
3.6. Inhibition of NF-kB by Notch-1 is not due to competition for the coactivator
CBP/p300 34
3.7. Regulatory effects of Notch-1-IC and Notch-2-IC on NF-kB activity are not correlated
with Hes-1 activation 36
3.8. Notch-1-IC decreases while Notch-2-IC activates NF-kB DNA-binding activity 38
3.9. Ectopic Notch-1-IC stabilizes whereas Notch-2-IC reduces IkBo protein levels after
LPS treatment 44
3.10. Expression profile of Traf-I and Traf-2 in Notch-1-IC and Notch-2-IC infectants .46
3.11. Increased downregulation of Toll-like receptor-4 surface expression in ectopic
Notch-2-IC-32D cells in response to LPS 49
I
3.12. Notch-1 -IC and Notch-2-IC appear neither to associate with p50 nor p65 subunit in
32D cells 52
3.13. Effect of ectopic Notch-1-IC and Notch-2-IC on the apoptosis in 32D cells after IL-3
withdrawal 55
4. Discussion 58
5. Outlook 66
6. Materials and Methods 67
6.1. Materials 67
6.1.1. Antibodies 67
6.1.2. Vectors 67
6.1.3. Oligonucleotides 68
6.2. Plasmids and Notch Constructs 68
6.3. DNA Cloning Procedures and DNA Sequencing 69
6.4. Cell Culture 69
6.5. Transient and Stable Transfections 70
6.5.1. Transfection of Helper Cells with Viral Constructs (Superfect Reagent) 70
6.5.2. Retroviral Transduction 71
6.5.3. Electroporation 71
6.5.4. Nucleoporation using the Amaxa System 72
6.6. Reporter Gene Assays 72
6.6.1. Luciferase Assay (Promega) 72
6.6.2. Dual Luciferase Assay (Promega) 73
6.7. Western Blot Analysis and Immunoprecipitation Studies 73
6.8. NF-kB Electrophoretic Mobility Shift Assay (EMSA) 75
6.9. NF-KBp65 Transcription Factor Assay (TransAM, Active Motif) 76
6.10. Function Elisa™ kBa Phosphorylation Assay (Active Motif) 77
6.11. Preparation of Cellular RNA and RT-PCR 77
6.12. FACS Analysis 79
6.12.1. Detection of Apoptosis (AnnexinV-FTTC and Yopro-iodide staining) 79
6.12.2. Toll-like receptor-4 extracellular Staining 80
7. Abbreviation Index 81
8. Literature 84
2 |
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| illustrated | Illustrated |
| indexdate | 2025-03-19T15:01:19Z |
| institution | BVB |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-012847640 |
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| physical | 96 S. Ill., graph. Darst. |
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| spelling | Dobierzewska, Aneta 1972- Verfasser (DE-588)129290351 aut Influence of notch dependent signaling on NF-kappaB activity and apoptosis of myelocytic cells von Aneta Dobierzewska 2004 96 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Erlangen-Nürnberg, Univ., Diss., 2004 Gen notch (DE-588)4762370-6 gnd rswk-swf Transkriptionsfaktor (DE-588)4303350-7 gnd rswk-swf Myelozyt (DE-588)4309927-0 gnd rswk-swf Signaltransduktion (DE-588)4318717-1 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Myelozyt (DE-588)4309927-0 s Transkriptionsfaktor (DE-588)4303350-7 s Gen notch (DE-588)4762370-6 s Signaltransduktion (DE-588)4318717-1 s DE-604 HBZ Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=012847640&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Dobierzewska, Aneta 1972- Influence of notch dependent signaling on NF-kappaB activity and apoptosis of myelocytic cells Gen notch (DE-588)4762370-6 gnd Transkriptionsfaktor (DE-588)4303350-7 gnd Myelozyt (DE-588)4309927-0 gnd Signaltransduktion (DE-588)4318717-1 gnd |
| subject_GND | (DE-588)4762370-6 (DE-588)4303350-7 (DE-588)4309927-0 (DE-588)4318717-1 (DE-588)4113937-9 |
| title | Influence of notch dependent signaling on NF-kappaB activity and apoptosis of myelocytic cells |
| title_auth | Influence of notch dependent signaling on NF-kappaB activity and apoptosis of myelocytic cells |
| title_exact_search | Influence of notch dependent signaling on NF-kappaB activity and apoptosis of myelocytic cells |
| title_full | Influence of notch dependent signaling on NF-kappaB activity and apoptosis of myelocytic cells von Aneta Dobierzewska |
| title_fullStr | Influence of notch dependent signaling on NF-kappaB activity and apoptosis of myelocytic cells von Aneta Dobierzewska |
| title_full_unstemmed | Influence of notch dependent signaling on NF-kappaB activity and apoptosis of myelocytic cells von Aneta Dobierzewska |
| title_short | Influence of notch dependent signaling on NF-kappaB activity and apoptosis of myelocytic cells |
| title_sort | influence of notch dependent signaling on nf kappab activity and apoptosis of myelocytic cells |
| topic | Gen notch (DE-588)4762370-6 gnd Transkriptionsfaktor (DE-588)4303350-7 gnd Myelozyt (DE-588)4309927-0 gnd Signaltransduktion (DE-588)4318717-1 gnd |
| topic_facet | Gen notch Transkriptionsfaktor Myelozyt Signaltransduktion Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=012847640&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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