Altered expression of pyruvate metabolism-related genes in <i>Npc1</i>^-/- cerebellum.

<p>Cerebellar tissue homogenates and RNA were prepared from 3-, 5-, or 7-week old wildtype (WT) and <i>Npc1</i>^-/- mice. A) Schematic representation of pathways of pyruvate metabolism showing mRNA (italics) and proteins analyzed in this study. The grey shading represents the mitochondrial compartment. B) mRNA levels of pyruvate dehydrogenase subunit A (<i>Pdha</i>) analyzed by qPCR standardized to <i>Ppia</i>. C) Immunoblots of tissue homogenates using antibodies against phospho-PDH and PDH, and actin as loading control. The bar graph shows the ratio of phospho-PDH per PDH, and PDH per actin as percent of the average of WT samples on the same membrane. D – L) mRNA levels of pyruvate dehydrogenase kinase 1 and 2 (<i>Pdk1</i> and <i>Pdk2</i>), pyruvate carboxylase (<i>Pcx</i>), cytosolic, mitochondrial, and neuronal mitochondrial malic enzymes (<i>Me1, Me2, Me3</i>), Aralar, Calbindin (<i>Calb1</i>), and neurofilament heavy polypeptide (<i>Nefh</i>) analyzed by qPCR standardized to <i>Ppia</i>. All bar graphs show the mean ± SEM. * p<0.05, <i>Npc1</i>^-/- vs. WT.