Extended examination of HD-associated mRNAs by qPCR from blood of HD monkeys. Five of the selected HD-associated mRNA transcripts with strong divergent trends (FZD8, NDUFA5, ALOX12, LMNA, LXN; panels A-E) were further subjected to evaluation by qPCR at extended developmental timespans of 29, 32, and 39 months of age from all control and HD monkeys. Additionally, the 5, 11, 17, and 23 month results timepoints for those same 5 candidates, initially examined by microarray, were also validated by qPCR. White blood cells were collected at each timepoint, followed by RNA extraction and real-time PCR (qPCR) analysis with gene specific Taqman assays (Applied Biosystems). The samples for qPCR were repeated in duplicates for each monkey. The values for all 4 controls were analyzed together and the values for all 4 HD monkeys were analyzed together; one blood sample was analyzed per monkey per timepoint. All qPCR results were normalized with the geometric mean of two endogenous controls, beta-actin (ACTB) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). A table of stats, including P values and FDRs for disease-association and age-effect by Two-Way ANOVA with repeated measures are outlined in panel F.
Kocerha et al. BMC Neuroscience 2013 14:88 doi:10.1186/1471-2202-14-88