Increased phosphofructokinase levels in <i>Npc1</i>^-/- cerebellum at the early symptomatic stage of the disease.

<p>A to C) Cerebellar tissue homogenates were prepared from 3- or 5-week old wildtype (WT) and <i>Npc1</i>^-/- mice and analyzed by immunoblotting using antibodies against hexokinase 1 (HK1; panel A), phosphofructokinase 1 (PFK1P; panel B), both isoforms 1 and 2 of pyruvate kinase type M (PKM1/2; panel C), and isoform 2 of PKM alone (PKM2, panel C) and actin as a loading control (all panels A to C). Bar graphs show the ratio of the band intensities of the proteins of interest to band intensity of actin immunoreactivity on the same membrane and are expressed as percent of the average of WT samples on the same membrane. D) Primary cortical neurons from embryonic WT and <i>Npc1</i>^-/- mice were incubated for 30 min in HEPES-buffered saline with [³H]-2-deoxyglucose and 2 mM glucose. Shown is the cell-associated radioactivity standardized to cell protein and expressed as percent of WT neurons of the same experiment (3 independent experiments in duplicate). All bar graphs show the mean ± SEM. * p<0.05, <i>Npc1</i>^-/- vs. WT.