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Series GSE8531 Query DataSets for GSE8531
Status Public on Aug 15, 2007
Title Profiling Motility Signal-Induced Genes in Human Keratinocytes
Organism Homo sapiens
Experiment type Expression profiling by array
Summary In an acute skin wound, newly released serum growth factors in the wound bed drive lateral migration of human keratinocytes (HKs) to re-epithelialize the wound. However, profiling the migration signal-specific genes has long been challenged by pleiotropic effects of a given growth factor, including proliferation, migration and factor-specific responses. To overcome these technical problems, we 1) took advantage of a unique response of HKs to transforming growth factor-beta (TGFbeta) to selectively suppress growth signal-responding genes and identify motility-specific genes and 2) employed dual stimulation of HKs with TGFalpha and insulin to identify the common genes and eliminate factor-specific genes. Under these conditions, DNA microarray analyses were utilized to study the profiles of both TGFalpha-regualted and insulin-regulated immediate early (IE, 30 min), early (E, 60 min) and delayed early (DE, 120 min) genes.
Keywords: time course, growth factor response
Overall design HKs were seeded on collagen I pre-coated tissue culture plates (150 mm) around 45% confluence (~ 2.5 X 106 cells) in complete medium and incubated overnight. Next day, the cells (~ 4.5 X 106 cells) were deprived of any supplemental GFs and incubated in serum-free medium for 16 hours to rest the cells at G0/G1 phase. The cells were then washed with pre-warmed serum-free medium and incubated in fresh serum-free medium containing 20 ng/ml TGFbeta for 20 min at 37oC to block expression of growth signal-related genes. TGFalpha or insulin was added to separate plates (in the continued presence of TGFbeta) for 30 min, 60 min and 120 min. At the end of each stimulation time points, the cells were washed with ice-cold PBS to holt the stimulation and subjected to RNA isolation.
Contributor(s) Cheng C, Fan J, Bandyopahdhay B, Guan S, Chen M, Woodley DT, Li W
Citation(s) 18323786
Submission date Jul 19, 2007
Last update date Dec 06, 2018
Phone 323-224-7057
Organization name USC
Lab CRL206
Street address 1303 N MISSION RD
State/province CA
ZIP/Postal code 90033
Country USA
Platforms (1)
GPL571 [HG-U133A_2] Affymetrix Human Genome U133A 2.0 Array
Samples (18)
GSM211154 30min control
GSM211157 30min TGFalpha
GSM211158 30min insulin
BioProject PRJNA101663

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Supplementary file Size Download File type/resource
GSE8531_RAW.tar 37.5 Mb (http)(custom) TAR (of CEL, CHP)
Processed data included within Sample table
Processed data provided as supplementary file

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