|
| Status |
Public on Feb 07, 2009 |
| Title |
Adjuvant mice with MLIF, biological rep2 |
| Sample type |
RNA |
| |
|
| Source name |
Articular tissue of mice treated with emulsified CFA, and MLIF, sacrified at four week after CII booster
|
| Organism |
Mus musculus |
| Characteristics |
strain: DBA1J
gender: Male
age: 8 weeks older
tissue: paw articular
|
| Treatment protocol |
CIA group, was immunized intradermally (i.d) at the base of the tail with 150 μg of Chicken collagen II (CII) (Sigma Chemical Co., St Louis MO, USA), dissolved in 5mM acetic acid and emulsified (1:1) in complete Freund adjuvant (CFA) (Sigma). According to established protocols, 3 weeks afterward, a booster using this time incomplete Freund adjuvant (IFA) was administered [9] CIA/MLIF-group animals; in addition to CIA protocol, 50 mg of 98 % pure MLIF (EzBiolab, Inc. Westfield, IN, USA) dissolved in 10 ml sterile and pyroxenes-free PBS was applied nasally weekly from 1 week prior to the first CII injection until the end of the experiment. CFA group was immunized i.d. at the base of the tail with 60ml of CFA followed by a boost of IFA 3 weeks latter. MLIF group was treated as CFA group in addition to a weekly nasal application of 50mg of MLIF during 9 weeks.
|
| Growth protocol |
DBA1/J male mice (Harlan México) maintained under standard conditions at the care facility and fed ad libitum. Mice aged 8-10 weeks and divided into four groups of 8 mice each, were employed.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Excised mice paws were triturated in Trizol using a politron PT-200 and homogenized into Qiashredder columns. Total RNA extraction was carried out from homogenized material by acid-phenol:chloroform extraction and further purified on a column (QIAGEN). Samples were run on a RNA 6000 Nano Gel System for RNA quality determination.
|
| Label |
biotin
|
| Label protocol |
Labeling protocol was carried out with 300 ng of purified RNA, with biotin, accord to Affymetrix Gene Mouse Arrays 1.0 ST protocol.
|
| |
|
| Hybridization protocol |
Following fragmentation, 5.5 mg of biotinilated RNA were hybridized for 16 hr at 45C into individual Affymetrix Gene Mouse Arrays 1.0 ST. GeneChips were washed and stained in the GeneChip® Fluidics Station 450 (Affymetrix) according to GeneChip® Whole Transcript (WT) Sense Target Labeling Assay manual (Affymetrix) instructions.
|
| Scan protocol |
Microarrays were scanned in a GeneChip® Scanner 3000 7G (Affymetrix) and quality control was evaluated by Affymetrix software (Expresion ConsoleTM).
|
| Description |
Adjuvant mice with MLIF, biological rep2
|
| Data processing |
Analysis of generated cell files for each microarray included Benjamini and Hochberg corrections. Normalized dates from Gene arrays were obtained by utilizing GeneSifterTM (VizX Labs LLC, Seattle, WA, USA) (www.genesifter.net). Our scrutiny included the 28,833 genes of the Mouse Gene Array by performing comparison of CFA vs CIA, CFA vs CIA/MLIF, CFA vs MLIF, and CIA vs CIA/MLIF mice groups.
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| |
|
| Submission date |
Feb 05, 2009 |
| Last update date |
Feb 06, 2009 |
| Contact name |
Juan Velazquez |
| E-mail(s) |
velazquez_juan@hotmail.com
|
| Organization name |
Instituto Mexicano del Seguro Social
|
| Street address |
Alameda 45
|
| City |
Zacatecas |
| State/province |
Zacatecas |
| ZIP/Postal code |
98000 |
| Country |
Mexico |
| |
|
| Platform ID |
GPL6246 |
| Series (1) |
| GSE14721 |
GENE EXPRESSION OF AN ANTI-INFLAMMATORY PENTAPEPTIDE ON CIA MURINE DEVELOPMENT |
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