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| Status |
Public on Dec 11, 2009 |
| Title |
KLF2-induced actin shear fibers control both alignment to flow and JNK signaling in vascular endothelium |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by array
Protein profiling by protein array
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| Summary |
The shear stress-induced transcription factor Krüppel-like factor 2 (KLF2) confers anti-inflammatory properties to endothelial cells through inhibition of activator protein 1, presumably by interfering with MAPK cascades. To gain insight into the regulation of these cascades by KLF2, we used antibody arrays in combination with time-course mRNA micro-array analysis. No gross changes in MAPKs were detected, rather phosphorylation of actin cytoskeleton-associated proteins, including Focal Adhesion Kinase, was markedly repressed by KLF2. Furthermore, we demonstrate that KLF2-mediated inhibition of Jun NH2-terminal kinase (JNK) and its downstream targets ATF2/c-Jun is dependent on the cytoskeleton. Specifically, KLF2 directs the formation of typical short basal actin filaments, we term shear fibers, which are distinct from thrombin- or TNF-α-induced stress fibers. KLF2 is shown to be essential for shear stress-induced cell alignment, concomitant shear fiber assembly and inhibition of JNK signaling. These findings link the specific effects of shear-induced KLF2 on endothelial morphology to the suppression of JNK MAPK signaling in vascular homeostasis via novel actin shear fibers.
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| Overall design |
Tramscriptome profiling: Three independent isolates of Human Umbilical Vein Endothelial cells were transduced with lentiviral vectors expressing Kruppel Like Factor 2 (KLF2) or no protein (mock), and at time after transduction 24 h, 48 h, 72 h , RNA was isolated and hybridized to GPL4868 microarrays using dye swap procedure
Kinome profiling: Two independent isolates of Human Umbilical Vein Endothelial cells were transduced with lentiviral vectors expressing Kruppel Like Factor 2 (KLF2) or no protein (mock), and at time after transduction 72 h , total cellular protein was isolated and hybridized to Kinexus KAM-1.1 phosphoprotein (kinexus) microarrays using dual color procedure in duplicate
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| Contributor(s) |
Horrevoets AJ, Boon RA |
| Citation(s) |
20032497 |
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| Submission date |
Dec 10, 2009 |
| Last update date |
Jan 18, 2013 |
| Contact name |
Anton J Horrevoets |
| URL |
http://www.vumc.nl/afdelingen/moleculairecelbiologie
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| Organization name |
VU University Medical Center
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| Department |
Molecular Cell Biology
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| Lab |
B244
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| Street address |
van der Boechorststraat 7
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| City |
Amsterdam |
| ZIP/Postal code |
1081 BT |
| Country |
Netherlands |
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| Platforms (2) |
| GPL4868 |
AMC-MAD Homo sapiens 19k ver 1.0 |
| GPL9787 |
KINEXUS KAM-1.1 kinome array |
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| Samples (26)
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| GSM483085 |
24 H KLF2 EXPRESSION,ISOLATE 1 |
| GSM483086 |
24 H KLF2 EXPRESSION,ISOLATE 1, REVERSE LABEL |
| GSM483087 |
24 H KLF2 EXPRESSION,ISOLATE 2 |
| GSM483088 |
24 H KLF2 EXPRESSION,ISOLATE 2, REVERSE LABEL |
| GSM483089 |
24 H KLF2 EXPRESSION,ISOLATE 3 |
| GSM483090 |
24 H KLF2 EXPRESSION,ISOLATE 3, REVERSE LABEL |
| GSM483091 |
48 H KLF2 EXPRESSION,ISOLATE 1 |
| GSM483092 |
48 H KLF2 EXPRESSION,ISOLATE 1, REVERSE LABEL |
| GSM483093 |
48 H KLF2 EXPRESSION,ISOLATE 2 |
| GSM483094 |
48 H KLF2 EXPRESSION,ISOLATE 2, REVERSE LABEL |
| GSM483095 |
48 H KLF2 EXPRESSION,ISOLATE 3 |
| GSM483096 |
48 H KLF2 EXPRESSION,ISOLATE 3, REVERSE LABEL |
| GSM483097 |
72 H KLF2 EXPRESSION,ISOLATE 1 |
| GSM483098 |
72 H KLF2 EXPRESSION,ISOLATE 1, REVERSE LABEL |
| GSM483099 |
72 H KLF2 EXPRESSION,ISOLATE 2 |
| GSM483100 |
72 H KLF2 EXPRESSION,ISOLATE 2, REVERSE LABEL |
| GSM483101 |
72 H KLF2 EXPRESSION,ISOLATE 3 |
| GSM483102 |
72 H KLF2 EXPRESSION,ISOLATE 3, REVERSE LABEL |
| GSM483212 |
mock-1 [kinome] |
| GSM483213 |
mock-2 [kinome] |
| GSM483214 |
mock-3 [kinome] |
| GSM483215 |
mock-4 [kinome] |
| GSM483216 |
KLF2-1 [kinome] |
| GSM483217 |
KLF2-2 [kinome] |
| GSM483218 |
KLF2-3 [kinome] |
| GSM483219 |
KLF2-4 [kinome] |
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| Relations |
| BioProject |
PRJNA121787 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE19412_kinome_KLF2_raw_OriginalData.txt.gz |
64.7 Kb |
(ftp)(http) |
TXT |
| GSE19412_kinome_KLF2k_raw_FilteredData.txt.gz |
65.7 Kb |
(ftp)(http) |
TXT |
| GSE19412_kinome_mock_raw_FilteredData.txt.gz |
64.7 Kb |
(ftp)(http) |
TXT |
| GSE19412_kinome_mock_raw_OriginalData.txt.gz |
63.7 Kb |
(ftp)(http) |
TXT |
| GSE19412_mRNAprofiling_rawdata.txt.gz |
1.7 Mb |
(ftp)(http) |
TXT |
| Processed data included within Sample table |
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