C57 b6 mice

Human GBM cell populations or GL261 cells were injected as previously described [29 (link)–31 (link)]. Four-week-old female wild-type C57B/6 mice were purchased from Charles River Laboratories and used between 4 and 5 weeks of age for all experiments. A total of 20,000 GL261 cells were resuspended in 20 μl of RPMI null media and injected intracranially into the left hemisphere 2 mm caudal to the coronal suture, 3 mm lateral to the sagittal suture at a 90° angle with the murine skull to a depth of 2.5 mm. Mice were monitored twice daily for signs of tumor burden and sacrificed when symptomatic or when tumor volume exceeded 5% of body weight. 5-fluorouracil (5-FU, Sigma) was prepared fresh for every treatment by resuspending in DMSO and diluted in PBS to achieve working concentrations. 5-FU or equivalent concentrations of DMSO were injected intraperitoneally on a weekly basis for all treatment arms of the in vivo experiments. Mice were imaged every 2–3 days by injection of luciferin (50 μl) intraperitoneally after anesthetization by isoflurane and imaged using an IVIS in vivo imaging system (Brucker) to monitor tumor growth. Survival curves were generated and analyzed using SigmaStat (version 3.5.0.54, Dundas Software).

All animal procedures were approved by the Duke University School of
Medicine Animal Care and Use Committee, Max Planck Florida Institute for
Neuroscience, and Weill Cornell Medical College Institutional Animal Care and
Use Committees and were conducted in accordance with the NIH Guide for the Care
and Use of Laboratory Animals.
We used both male and female rats and mice. Rats and C57/B6 mice were
obtained from Charles River, Trkb^F616A mutant mice
were provided by D. Ginty^{28 (link)},
Bdnf^fl/fl and
Trkb^fl/fl mice were provided by L.
Parada^{32 (link)}, and
Bdnf-HA mice were generated as previously
described^{26 (link)}. The
genotype of each animal used was verified before and after preparing slices
using PCR of genomic DNA isolated from tail DNA before and slice samples
after.