In MP experiment, animals were randomly assigned into four groups: spinal cord injury with 70% ethanol (SCI + EtOH), spinal cord injury with MP (30 mg/kg) (SCI + MP), Sham + EtOH and Sham + MP. In EPO experiment, animals were randomly assigned into four groups: spinal cord injury with H2O (SCI + H2O), spinal cord injury with EPO (5000 U/kg) (SCI + EPO), Sham + H2O and Sham + EPO. Animals were sacrificed at 2, 4, 6, 8, 12, 24 and 48 hours after medication. Additional animals including Sham + EtOH, SCI + EtOH, Sham + H2O, and SCI + H2O were sacrificed immediately after control medication. For all surgeries, animals were anesthetized by a mixture of Zoletil 50 (25 mg/kg) and Rompun (xylazine, 10 mg/Kg) via intraperitoneal injections.
Growth protocol
Long Evans female rats (8 or 9 week-old) were purchased from National Laboratory Animal Center, Taipei, Taiwan. All animals were housed in a 12-h light/dark cycle and had free access to water and food all the time. For animal care and surgical procedures, we followed the National Institutes of Health Guide for the Care and Use of Laboratory Animals (NIH Publications No. 80-23, revised 1978) and all surgical protocols were approved by the Animal Care and Use Committee of Taipei Medical University.
Extracted molecule
total RNA
Extraction protocol
Following different time after MP, EPO or control medication, SCI animals were anesthetized with a mixture of Zoletil 50 and Rompun described earlier and the 5-mm lesion spinal cord was removed, frozen immediately in liquid nitrogen and stored in liquid nitrogen prior to RNA extraction. Total RNA was extracted by the Trizol method and the integrity of RNA was examined by Agilent Bioanalyzer (Agilent Technologies, Inc. Headquarters, Palo Alto, CA, USA) before oligo microarray analysis started.
Label
biotin
Label protocol
According to the Affymetrix manufactor's manual.
Hybridization protocol
According to the Affymetrix manufactor's manual.
Scan protocol
According to the Affymetrix manufactor's manual.
Description
Gene Expression data
Data processing
The data are normalized using RMA in GeneSpring GX 10.
