|
| Status |
Public on Dec 13, 2011 |
| Title |
High dosage of LECT2 vs control, 48 hours, biological replicate 2 of 2 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
Mouse peritoneal macrophages exposed to 0 μg/ml of LECT2 for 48 hours
|
| Organism |
Mus musculus |
| Characteristics |
tissue: mouse peritonaeum
treatment: exposed to 0 μg/ml of LECT2 for 48 hours
|
| Treatment protocol |
Mouse peritoneal macrophages were plated at 1×105 cells/cm2 in complete culture medium and incubated with 0,0.5 or 5 μg/ml LECT2 for 3.5 or 48 hours
|
| Growth protocol |
Mouse peritoneal macrophages were isolated from mouse peritonaeum. After washed with phosphate-buffered saline (PBS) twice, peritoneal macrophages were plated and cultivated in Dulbecco’s modified Eagle’s medium-low glucose supplemented with 20% FBS.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA from different cultured cells was extracted using Trizol (Invitrogen) according to the manufacturer’s instructions.
|
| Label |
Cy5
|
| Label protocol |
1μg of total RNA was used to synthesize the double strand cDNA. RNA was amplified by in vitro transcription using Ambion’s MessageAmp™ II aRNA Amplification Kits (Life Technologies, Austin, TX, USA). Then, aRNA was reverse transcribed into cDNA and further labeled with cy3-dCTP with Klenow enzyme.
|
| |
|
| Channel 2 |
| Source name |
Mouse peritoneal macrophages exposed to 5 μg/ml of LECT2 for 48 hours
|
| Organism |
Mus musculus |
| Characteristics |
tissue: mouse peritonaeum
treatment: exposed to 5 μg/ml of LECT2 for 48 hours
|
| Treatment protocol |
Mouse peritoneal macrophages were plated at 1×105 cells/cm2 in complete culture medium and incubated with 0,0.5 or 5 μg/ml LECT2 for 3.5 or 48 hours
|
| Growth protocol |
Mouse peritoneal macrophages were isolated from mouse peritonaeum. After washed with phosphate-buffered saline (PBS) twice, peritoneal macrophages were plated and cultivated in Dulbecco’s modified Eagle’s medium-low glucose supplemented with 20% FBS.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA from different cultured cells was extracted using Trizol (Invitrogen) according to the manufacturer’s instructions.
|
| Label |
Cy3
|
| Label protocol |
1μg of total RNA was used to synthesize the double strand cDNA. RNA was amplified by in vitro transcription using Ambion’s MessageAmp™ II aRNA Amplification Kits (Life Technologies, Austin, TX, USA). Then, aRNA was reverse transcribed into cDNA and further labeled with cy3-dCTP with Klenow enzyme.
|
| |
|
| |
| Hybridization protocol |
According to the instructions of Agilent Gene Expression Hybridization Kit
|
| Scan protocol |
Scanned on an Agilent G2505C scanner.Images were quantified using Agilent Feature Extraction Software (version 10.7.3.1).
|
| Description |
Biological replicate 2 of 2, 5 μg/ml of LECT2 for 48 hours vs 0 μg/ml of LECT2 for 48 hours
|
| Data processing |
Agilent Feature Extraction Software (version 10.7.3.1) was used for background subtraction and LOWESS normalization.
|
| |
|
| Submission date |
Nov 15, 2011 |
| Last update date |
Dec 13, 2011 |
| Contact name |
wang liang |
| E-mail(s) |
[email protected]
|
| Organization name |
BioChainBJ
|
| Street address |
7A North Yongchang Road BDA
|
| City |
Beijing |
| ZIP/Postal code |
100176 |
| Country |
China |
| |
|
| Platform ID |
GPL11202 |
| Series (1) |
| GSE33721 |
Mouse peritoneal macrophages at 3.5 or 48 hours, 0,0.5 or 5 μg/ml Leukocyte cell-derived chemotaxin 2 (LECT2),gene expression array, two independent biological replicates |
|
