|
| Status |
Public on Apr 16, 2013 |
| Title |
TCDD treated Hepa1c1c7 vs. TCDD plus blebbistatin |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
Hepa1c1c7 cells treated with TCDD
|
| Organism |
Mus musculus |
| Characteristics |
cell line: Hepa1c1c7 (ATCC: CRL-2026)
days after passage: 4 days after passage
tissue: liver, carcinoma
strain: C57BL/6
treatment: TCDD
|
| Growth protocol |
Hepa1c1c7 cells were cultured in DMEM medium containing 0.1 U/L penicillin, 0.1 g/L streptomycin and 5% fetal bovine albumin.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extracted using Rneasy Kit (QIAGEN) following manufacturer's instructions.
|
| Label |
Cy3
|
| Label protocol |
200 ng of total RNA were primed with 1.8 µl T7 Promoter Primer at 65°C for 5 min, then reversed transcribed at 40°C for 2 h in the presence of 1.2 uL AffinityScript, and 100 µM each dATP, dTTP, dGTP, with 25 µM dCTP, 25 µM Cy3 or Cy5 (Agilent Technologies).
|
| |
|
| Channel 2 |
| Source name |
Hepa1c1c7 treated with TCDD plus blebbistatin
|
| Organism |
Mus musculus |
| Characteristics |
cell line: Hepa1c1c7 (ATCC: CRL-2026)
days after passage: 4 days after passage
tissue: liver, carcinoma
strain: C57BL/6
treatment: TCDD plus blebbistatin
|
| Growth protocol |
Hepa1c1c7 cells were cultured in DMEM medium containing 0.1 U/L penicillin, 0.1 g/L streptomycin and 5% fetal bovine albumin.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extracted using Rneasy Kit (QIAGEN) following manufacturer's instructions.
|
| Label |
Cy5
|
| Label protocol |
200 ng of total RNA were primed with 1.8 µl T7 Promoter Primer at 65°C for 5 min, then reversed transcribed at 40°C for 2 h in the presence of 1.2 uL AffinityScript, and 100 µM each dATP, dTTP, dGTP, with 25 µM dCTP, 25 µM Cy3 or Cy5 (Agilent Technologies).
|
| |
|
| |
| Hybridization protocol |
Oligoarray control targets and hybridization buffer (Agilent Hybridization Kit) were added, and samples were applied to microarrays enclosed in Agilent SureHyb-enabled hybridization chambers. After hybridization, slides were washed sequentially.
|
| Scan protocol |
Scanned on an Agilent Technologies Microarray scanner.
Images were quantified using Agilent Feature Extraction Software (version A.8.5.1.1).
|
| Description |
Hepa1c1c7 cells were treated with (-)-blebbistatin (40 uM) for 1 day and then TCDD (4 nM) plus blebbistatin for 6 h.
|
| Data processing |
Agilent Feature Extraction Software was used for background subtraction and LOWESS normalization.
|
| |
|
| Submission date |
Apr 15, 2013 |
| Last update date |
Apr 16, 2013 |
| Contact name |
Hideaki Kikuchi |
| E-mail(s) |
hkikuchi@cc.hirosaki-u.ac.jp
|
| Phone |
+81-172-39-3568
|
| Fax |
+81-172-39-3568
|
| Organization name |
Hirossaki University
|
| Department |
Faculty of Agriculture and Life Science
|
| Lab |
Biochemistry and Molecular Biology
|
| Street address |
3 Bunkyo-cho
|
| City |
Hirosaki |
| ZIP/Postal code |
036-8561 |
| Country |
Japan |
| |
|
| Platform ID |
GPL10333 |
| Series (1) |
| GSE46061 |
Mouse Hepa1c1c7 cells: TCDD vs. TCDD plus blebbistatin |
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