Extended Data Fig. 5: Experiments to evaluate mechanism of enhanced pseudouridylation by U > Ψ snRNAs.
From: Enhancing RNA base editing on mammalian transcripts with small nuclear RNAs
a, Rolling circle amplification FISH (RCA FISH) schematic, representative images and plots of rolonies per cell, mean rolony distance to nucleus/nucleolus and nucleolar-to-cellular rolony ratio for EEF2-targeting H/ACA box snoRNA and U > Ψ snRNAs. RCA FISH was performed in human U-2 OS cells with ×10 magnification images. Images depicting fluorescent signal are small representations of total number of nuclei (thousands per biological replicate) and are not suitable for visual interpretation. Scale bar: 20 microns. Pixel resolution: 752 nm x 752 nm. b, Guide-specific qPCR to quantitate expression levels of H/ACA box snoRNA and U > Ψ snRNA constructs targeting CFTR, ACTB, EEF2 and RPS6. Significance of difference: ***: p < 1e-3 (one-way ANOVA) (CFTR 3e-4, ACTB 3e-4). Error bars reflect standard error of mean. n = 3 biological replicates per condition.
