|
| Status |
Public on Aug 30, 2016 |
| Title |
500µM HC 0µM vesicant 8hr replicate 2 |
| Sample type |
RNA |
| |
|
| Source name |
Keratinocyte 24hr pretreatment with 500µM hydrocortisone (500µM HC) followed by 8hr incubation
|
| Organism |
Homo sapiens |
| Characteristics |
tissue: Normal Human Epidermal Keratinocytes (NHEK)
|
| Biomaterial provider |
Lonza (Walkersville, MD)
|
| Treatment protocol |
Five days after plating, cells were pre-treated with control vehicle (1.25% ethanol) or hydrocortisone (500µM HC). After 24 hours cells were exposed to either 200µM SM or 200µM NM for 1 hour or 8 hours. Control unexposed cells were included with the study.
|
| Growth protocol |
NHEK (Lonza, Walkersville, MD) were seeded at a density of 2.5X10E4 per well in T25 flasks and maintained according to manufucturer's specifications.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Following SM and NM exposure cells were gently rinsed twice with hanks buffered saline solution. Total RNA was isolated from exposed and control cells using miRNeasy Mini Kit (Qiagen, Valencia, CA)
|
| Label |
biotin
|
| Label protocol |
500ng total RNA was labeled using the RNA biotin labeling flashtag biotin HSR RNA labeling kit (Affymetrix, Santa Clara, CA) as per manufacturer’s instructions.
|
| |
|
| Hybridization protocol |
Labelled RNA was hybridized for 16 hrs at 48 degrees C on miRNA 3.0 GeneChips (Affymetrix). GeneChips were washed and stained using the Affymetrix Fluidics Station 450.
|
| Scan protocol |
GeneChips were scanned using Affymetrix 7G GeneChip Scanner.
|
| Description |
Cells maintained in humidified incubator at 37 degrees C and 5% CO during treatment
|
| Data processing |
Raw data (CEL files) were imported into Partek Genomic suite v6.6 (Partek Incorporated, St. Louis, MO) and normalized using robust multiarray averaging (RMA)
|
| |
|
| Submission date |
Feb 03, 2014 |
| Last update date |
Aug 30, 2016 |
| Contact name |
E Chepchumba K Yego |
| E-mail(s) |
eky1@case.edu
|
| Phone |
216-375-7334
|
| Organization name |
US Army Medical Research Institute of Chemical Defense
|
| Department |
Research
|
| Street address |
3100 Ricketts Point Road
|
| City |
Aberdeen Proving Ground |
| State/province |
MD |
| ZIP/Postal code |
21010 |
| Country |
USA |
| |
|
| Platform ID |
GPL16384 |
| Series (1) |
| GSE54639 |
microRNA (miRNA) expression data from normal human epidermal keratinocytes exposed to sulfur mustard and nitrogen mustard in the presence and absence of hydrocortisone treatment |
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