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Sample GSM566706 Query DataSets for GSM566706
Status Public on Jan 03, 2012
Title Cortical neurons-Lact-24h-Rep 1
Sample type RNA
Source name Murine primary cortical neurons
Organism Mus musculus
Characteristics strain: Swiss Albino mice
age: Gestation day 15-16
tissue: Primary cortical neurons
age of culture: Day 5
time point: 24h
agent: lactacystin
Treatment protocol Lactacystin was prepared as 1mM stock solution in DMSO stored at -20°C. Desired concentrations were achieved by dilution with serum-free NB. On day 5 in vitro, the cultured neurons were treated with 1uM lactacystin in NB medium.
Growth protocol Neocortical neurons (gestational days 15 or 16) obtained from foetal cortices of Swiss albino mice were used to prepare the primary cultures employing previous described procedures with modifications [Cheung NS, Beart PM, Pascoe CJ, John CA, Bernard O. Human Bcl-2 protects against AMPA receptor-mediated apoptosis. J Neurochem. 2000; 74: 1613-1620.].
Extracted molecule total RNA
Extraction protocol RNA from samples was extracted using RNeasy Mini Kit (Qiagen Cat. No. 74104) according to the manufacturer’s instructions. 1.5μl of the RNA sample was aliquoted for spectrophotometric quantification using Nanodrop ND-1000 Version 3.2.1 and 1μl for RNA quality analysis using E-gene HDAGT12 genetic analyzer.
Label biotin
Label protocol According to technical manual form Affymetrix, 7μg of extracted total RNA was used for cDNA synthesis. Double-stranded cDNA was synthesized using Superscript II reverse transcriptase (Invitrogen, Carlsbad, CA) with a T7-dT24 primer. After cleanup, Biotin-labeled cRNA was synthesized by in vitro transcription (Enzo Diagnostic, Inc., NY, USA) and fragmented subsequently.
Hybridization protocol 15μg of fragmented cRNA produced above was hybridized to the GeneChip Murine genome U74A and U74Av2 arrays for 16h at 45°C. GeneChips were washed and stained in the Affymetrix Fluidics Station 400.
Scan protocol The hybridized arrays were washed, stained, and scanned using the Hewlett-Packard GeneArray Scanner G2500A according to the manufacturer’s instructions.
Description Replicate 1
Data processing The data from each array were collected and initially analyzed using Affymetrix Microarray Suite 5.0 software. For comparison of multiple arrays, the signal intensity of each array was scaled to 500. The regulated genes were filtered on fold change 1.5 fold against controls in at least one of three time-points. One-way ANOVA (p<0.05) approach was used to find differentially expressed genes using GeneSpring™ GX 7.3 software (Agilent Technologies, CA, USA).
Submission date Jul 15, 2010
Last update date Jan 03, 2012
Contact name Minghui Jessica Chen
Organization name Menzies Research Institute
Department Neuroscience group
Lab A/P Steve Cheung
Street address Menzies Research Institute, University of Tasmania, Private Bag 24
City Hobart
State/province Tasmania
ZIP/Postal code 7001
Country Australia
Platform ID GPL32
Series (1)
GSE22465 Global transcriptomic profiling of lactacystin-mediated neuronal death

Data table header descriptions
VALUE MAS5.0 signal intensity

Data table
AFFX-MurIL2_at 13.1 A 0.814869
AFFX-MurIL10_at 24.1 A 0.5
AFFX-MurIL4_at 20.6 A 0.250796
AFFX-MurFAS_at 164.1 P 0.006187
AFFX-BioB-5_at 456.6 P 0.001102
AFFX-BioB-M_at 753.2 P 0.000044
AFFX-BioB-3_at 495.3 P 0.000095
AFFX-BioC-5_at 974.1 P 0.00011
AFFX-BioC-3_at 827.8 P 0.000044
AFFX-BioDn-5_at 1387 P 0.000044
AFFX-BioDn-3_at 6134 P 0.000081
AFFX-CreX-5_at 13042 P 0.000044
AFFX-CreX-3_at 16691.5 P 0.000044
AFFX-BioB-5_st 13.9 A 0.574038
AFFX-BioB-M_st 51.8 A 0.368438
AFFX-BioB-3_st 20 A 0.659339
AFFX-BioC-5_st 9.3 A 0.916408
AFFX-BioC-3_st 5.1 A 0.772364
AFFX-BioDn-5_st 150.5 A 0.078002
AFFX-BioDn-3_st 137.9 M 0.046482

Total number of rows: 10043

Table truncated, full table size 256 Kbytes.

Supplementary file Size Download File type/resource
GSM566706.CEL.gz 2.8 Mb (ftp)(http) CEL
Processed data included within Sample table

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