Tissue tek oct compound

Snap-frozen and OTC-embedded (Tissue-Tek® OCT Compound, Ted
Pella, INC., Redding, CA) of 15 GBM specimens were sectioned using a Microm
HM525 NX Cryostat (Thermo Fisher Scientific Inc., Grand Island, NY); one of
many serial consecutive sections per case was stained with hematoxylin and
eosin to confirm the localizations of tumor cells in different regions.
Tumor cells in different regions were subsequently isolated by laser-capture
microdissection using a Zeiss PALM (Carl Zeiss, Inc., Thornwood, NY). For
each case, 3 regions of tumor cells were separately collected for RNA
extraction. Total RNA was isolated from the cells using TRIzol RNA Isolation
Reagent according to the manufacturer’s instructions. The
concentrations and quality of the recovered RNA were measured using a
NanoDrop 2000 spectrophotometer (Nanodrop Technologies, Wilmington, DE). For
quantitative analysis of mRNA expressions of FOXM1-AS and FOXM1 by qPCR,
mRNA levels were normalized to GAPDH mRNA.

Mice were anesthetized and intracardially perfused first with 10 ml of PBS and then with 20 ml of 4% paraformaldehyde in PBS. Spinal cords were dissected, postfixed in 4% paraformaldehyde in PBS for 1h, and then placed in a 30% sucrose solution in PBS overnight. Spinal cord samples were embedded in Tissue-Tek OCT compound (Ted Pella, Inc., Redding, CA), frozen on dry ice, and sectioned at 8μm with a cryostat. Cross-sections were used to visualize myelin gene expression within spinal cord myelinated tracts.