Figure 8.

Validation of RNA integrity on Fresh frozen NHP (MJ 61) frontal cortex brain tissue using RNAscope assay. NHP tissue was stained with a 3-plex positive control probe POLR2A targeting DNA-directed RNA polymerase II RPB1, PPIB targeting cyclophilin B, and UBC that targets Ubiquitin C. In terms of relative expression levels, UBC is highest, PPIB is considered a moderate-high, POLR2A is moderate-to low expressor target. Successful staining has 4–9 dots per cell of PPIB/POLR2A and UBC of 10–15 dots per cell. PPIB is considered a standard to determine the RNA quality of brain samples. (C) Represents red fluorescence, indicating the specificity of PPIB, whereas, (D) displays a far-red puncta targeting UBC gene. Absence of POLR2A signal in (B) represents, quality of RNA being partially compromised. A merge image of all the channels can be seen in (A). White arrows were pointing toward the fluorescence puncta and orange arrows toward autofluorescence of the tissue.