All of the animal experiments were conducted using protocols approved by the Institutional Animal Care and Use Committees of Fred Hutchinson Cancer Research Center, protocol 50842. Ntva-Ink4a/Arf+/- and Ntva-Ink4a/Arf-/- mice were used to generate mouse gliomas. The genetic backgrounds of tva mice are FVB/N, C57BL6, BALB/C, and 129. To generate mCherry and HoxA5-expressing mouse gliomas, we used the RCAS/tva system. Briefly, DF1 cells were maintained with 10% fetal bovine serum (FBS) in Dulbecco's Modified Eagle Medium (DMEM). Chicken fibroblasts were transfected with each RCAS viral plasmid using Fugene 6 Transfection reagent (Roche) following the manufacturer’s protocol. Then, PDGFb-expressing DF1 cells were mixed with either mCherry- or HoxA5-expressing DF1 cells. This mixture of DF1 cells was injected to Ntva-Ink4a/Arf+/- and Ntva- Ink4a/Arf-/- mice. Adult mice anesthetized with isoflurane were positioned in a stereotactic device (Stoelting, Wood Dale, IL). RCAS transfected DF1 cells were injected using a 30-gauge needle inserted into the right frontal cortex (coordinates: anterior to bregma 1.5 mm, lateral 0.5 mm, and a depth 1.5 mm). Then, mice were monitored daily until they developed signs of illness, such as lethargy, poor grooming, weight loss, dehydration, macrocephaly, seizure, jumping and paralysis.
Extracted molecule
total RNA
Extraction protocol
Total RNA was extracted from tumor tissues using RNeasy Mini kit (QIAGEN) using the manufacturer’s protocol.
Label
biotin
Label protocol
Standard Illumina hybridization protocolHoxA5 is the experimental group and the mCherry is the control group.
Hybridization protocol
Standard Illumina hybridization protocol
Scan protocol
Standard Illumina scanning protocol
Description
Sample 1 HoxA5 tumor in mice replicate 1
Data processing
The data were normalized using R/Bioconductor package limma
