Figure 8.

Pyruvate metabolism is reduced in PrP KO NSCs.A, LDH activity assay of WT and KO NSCs with (B) LDHA and LDHB gene expression. C, changes in the pyruvate dehydrogenase complex genes and pyruvate carboxylase transcripts associated with PrP KO in NSCs with the RNA-seq FPKM shown on the left y-axis and the qRT-PCR Delta Ct shown on the right y-axis for comparison. RNA samples were harvested for RNAseq and qRT-PCR independently. D, schematic showing the enzymes that catalyze pyruvate with those shown in red reduced and those shown in green increased in PrP KO NSCs. Graph in panel A is displayed relative to the average LDH activity of WT (n = 3). Results in A were compared by two-tailed unpaired student’s t test with Welch’s correction, ∗∗p < 0.01. For B and C, fold changes and p-values appropriate to the assay (see methods) are shown with each plot. Explanation of abbreviations can be found in Table 2. FPKM, Fragments per kilobase million; LDH, lactate dehydrogenase; NSC, neural stem cell; PrP, prion protein; qRT-PCR, quantitative reverse-transcription polymerase chain reaction.