Figure 5

Conditional deletion of Hk2 prevents Shh-induced aerobic glycolysis. (A) Representative Western blot comparing Hk2, Hk1 and Cyclin D2 expression in cerebellar lysates at P5 from Hk2^fl/fl and Hk2^+/+ mice with and without hGFAP-cre. Equivalent results were obtained in 5 pups for each genotype at each age. (B) EdU incorporation (green) 24 hours after IP injection demonstrates proliferation in the EGL of Hk2^fl/fl mice without cre (control) or of hGFAP-cre;Hk2^fl/fl mice. White arrowhead marks focus of ectopic proliferation. Nuclei are counterstained with DAPI. Scale bars = 100 μm. (C) Lactate accumulation in media from hGFAP-cre;Hk2^fl/fl CGNPs was markedly reduced compared with CGNPs of Hk2^fl/fl mice without cre. Data compiled from 3 replicates per condition and mean ± SEM values are presented as fold-change relative to control without cre. Two-tailed Student’s t test was used for statistical comparison. (D) Representative NMR spectra from media samples of Hk2^fl/fl mice without cre (control) or of hGFAP-cre;Hk2^fl/fl mice. Experiment performed with 3 replicates and mean ± SEM values presented. Lactate was the only detectable metabolite to be significantly altered by Hk2 deletion (P < 0.02, two-way analysis of variance with Bonferroni correction).