|
| Status |
Public on May 20, 2016 |
| Title |
Primary Rat_SQ1 treated_Replicate5 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
primary hepatocytes_SQ1
|
| Organism |
Rattus norvegicus |
| Characteristics |
strain: Sprague Dawley
gender: male
cell type: Primary cultured Rat Hepatocytes
treated with: SQ1
|
| Treatment protocol |
48 hours after plating, hepatocytes were left untreated or treated with 100 µM SQ1 or 30 µM Pravastatin for 48 hours.
|
| Growth protocol |
Primary hepatocytes plated at density and maintained in Williams Medium E.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted using Qiagen lysis buffer following manufacturer's instructions
|
| Label |
A647
|
| Label protocol |
500ng of total RNA was input to the TargetAMP 1-Round Aminoallyl-aRNA Amplification Kit 101 (Epicentre, Madison, WI) to produce aminoallyl-aRNA according to the vendor’s protocol. Five ug of each aminoallyl-aRNA sample was labeled with Alexa 555 or Alexa 647 (Molecular probes/ Life Technologies, Foster City, CA).
|
| |
|
| Channel 2 |
| Source name |
primary hepatocytes_untreated control
|
| Organism |
Rattus norvegicus |
| Characteristics |
strain: Sprague Dawley
gender: male
cell type: Primary cultured Rat Hepatocytes
treated with: none (untreated control)
|
| Treatment protocol |
48 hours after plating, hepatocytes were left untreated or treated with 100 µM SQ1 or 30 µM Pravastatin for 48 hours.
|
| Growth protocol |
Primary hepatocytes plated at density and maintained in Williams Medium E.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted using Qiagen lysis buffer following manufacturer's instructions
|
| Label |
A555
|
| Label protocol |
500ng of total RNA was input to the TargetAMP 1-Round Aminoallyl-aRNA Amplification Kit 101 (Epicentre, Madison, WI) to produce aminoallyl-aRNA according to the vendor’s protocol. Five ug of each aminoallyl-aRNA sample was labeled with Alexa 555 or Alexa 647 (Molecular probes/ Life Technologies, Foster City, CA).
|
| |
|
| |
| Hybridization protocol |
0.825µg of Alexa 555 labeled Aminoallyl-aRNA and 0.825µg of Alexa 647 labeled Aminoallyl-aRNA were mixed together and allowed to co-hybridize on the array for 17 hours at 65°C at 10rpm in the hybridization rotator. After hybridization the slide was washed with Agilent GE Wash Buffers following the Agilent’s protocol.
|
| Scan protocol |
Scanned on an Agilent G2565AA scanner. Images were quantified using Agilent Feature Extraction Software (version A.8.5.1.1).
|
| Description |
Rt SQ1_5
|
| Data processing |
Agilent Feature Extraction Software was used for background subtraction and LOWESS normalization.
|
| |
|
| Submission date |
May 19, 2016 |
| Last update date |
May 20, 2016 |
| Contact name |
Thomas Kocarek |
| E-mail(s) |
[email protected]
|
| Phone |
313-577-6580
|
| Organization name |
Wayne State University
|
| Department |
Institute of Environmental Health Sciences
|
| Street address |
6135 Woodward Ave
|
| City |
Detroit |
| State/province |
MI |
| ZIP/Postal code |
48202 |
| Country |
USA |
| |
|
| Platform ID |
GPL14746 |
| Series (2) |
| GSE81657 |
Rat Primary Cultured Hepatocytes: Control vs SQ1 or Pravastatin treatment. |
| GSE81659 |
Expression data from squalestatin 1- or pravastatin-treated primary cultured mouse and rat hepatocytes. |
|
