|
| Status |
Public on Sep 30, 2009 |
| Title |
BDK established from donor I_DNCB treated_4.5 hr |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
bulge-derived keratinocytes established from donor I_DNCB treated_4.5 hr
|
| Organism |
Homo sapiens |
| Characteristics |
gender: male
|
| Treatment protocol |
BDKs or NHEK ; seeded at 5 x 105 cells/60 mm φ dish and incubated in medium at 37 °C for 24 hr, and sequentially in MCDB153 medium (Research Institute for the Functional Peptides Co., Ltd, Yamagata, Japan) at 37 °C for 24 hr. 24 hr later, cells were treated with 10 μM DNCB or 0.1% DMSO for 4.5 hr. THP-1 ; seeded at the density of 7 x 105 cells/10 mm φ dish. 24 hr later, cells were treated with 10 μM DNCB or 0.1% DMSO for 4.5 hr.
|
| Growth protocol |
BDKs and NHEK ; cultured in Defined Keratinocyte-SFM medium （DK-SFM, GIBCO/Invitrogen Co., CA） at 37°C in a humidified atmosphere containing 5% CO2. THP-1; cultured in RPMI1640 medium (GIBCO/Invitrogen Co.) containing 10% fetal bovine serum (Equitech-bio Inc., TX, USA) at 37°C in a humidified atmosphere containing 5% CO2.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Isolated using TRIzol reagent (Invitrogen, CA, USA) and purified using an RNeasy kit (QIAGEN, Hilden, Germany).
|
| Label |
Cy5
|
| Label protocol |
Total RNAs (each 200ng) were labeled with Cy3- or Cy5- CTP using Agilent Quick Amp Labeling Kit (Agilent Technologies Inc.).
|
| |
|
| Channel 2 |
| Source name |
bulge-derived keratinocytes established from donor I_DMSO (solvent control) treated_4.5 hr
|
| Organism |
Homo sapiens |
| Characteristics |
gender: male
|
| Treatment protocol |
BDKs or NHEK ; seeded at 5 x 105 cells/60 mm φ dish and incubated in medium at 37 °C for 24 hr, and sequentially in MCDB153 medium (Research Institute for the Functional Peptides Co., Ltd, Yamagata, Japan) at 37 °C for 24 hr. 24 hr later, cells were treated with 10 μM DNCB or 0.1% DMSO for 4.5 hr. THP-1 ; seeded at the density of 7 x 105 cells/10 mm φ dish. 24 hr later, cells were treated with 10 μM DNCB or 0.1% DMSO for 4.5 hr.
|
| Growth protocol |
BDKs and NHEK ; cultured in Defined Keratinocyte-SFM medium （DK-SFM, GIBCO/Invitrogen Co., CA） at 37°C in a humidified atmosphere containing 5% CO2. THP-1; cultured in RPMI1640 medium (GIBCO/Invitrogen Co.) containing 10% fetal bovine serum (Equitech-bio Inc., TX, USA) at 37°C in a humidified atmosphere containing 5% CO2.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Isolated using TRIzol reagent (Invitrogen, CA, USA) and purified using an RNeasy kit (QIAGEN, Hilden, Germany).
|
| Label |
Cy3
|
| Label protocol |
Total RNAs (each 200ng) were labeled with Cy3- or Cy5- CTP using Agilent Quick Amp Labeling Kit (Agilent Technologies Inc.).
|
| |
|
| |
| Hybridization protocol |
Cy3- or Cy5-labeled probes were mixed, fragmented, and then dissolved in GE hybridization buffer in the Gene Expression Hybridization Kit (Agilent). After hybridization, slides were washed sequential.
|
| Scan protocol |
Scanned on an Agilent microarray scanner.
Images were quantified using Agilent Feature Extraction Software (ver9.5.3.1).
|
| Description |
BDKs ; prepared from plucked hairs using a depilation forceps from the side area of the scalp of adult subjects, who gave written informed consent
|
| Data processing |
LOWESS normalized, background subtracted data obtained from log2 of processed Red signal/processed Green signal. Expressionist (GeneData AG, Basel, Switzerland) software was used.
|
| |
|
| Submission date |
Feb 13, 2009 |
| Last update date |
Jul 26, 2009 |
| Contact name |
Yoshie Yoshikawa |
| E-mail(s) |
yoshiey@hyo-med.ac.jp
|
| Phone |
+81-798-45-6587
|
| Organization name |
Hyogo College of Medicine
|
| Department |
Genetics
|
| Street address |
1-1 Mukogawa-cho
|
| City |
Nishinomiya |
| State/province |
Hyogo |
| ZIP/Postal code |
663-8501 |
| Country |
Japan |
| |
|
| Platform ID |
GPL6480 |
| Series (1) |
| GSE14838 |
Gene expression changes by 2,4-dinitrochlorobenzene in human bulge-derived keratinocytes |
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