Fig. 4.

Reduction of scaRNA2 increases 28S rRNA G3923 methylation. (A) Complimentary alignment of snord111A/snord111B with 28S rRNA. Also shown is the base pairing of regRNP2 with snord111A/B. (B) RNA isolated from cells treated with control 1 or snord111B ASO (left panel), or control 2 or scaRNA2 siRNA (right panel), was subjected to primer extension with low (5 uM) dNTP levels. The location of the G3923 methylation stop is indicated, as is a band used for normalization. Quantification of this and other data are shown in C. Relative to control 1, reduction of the snord111B guide RNA decreases G3923 methylation by 71% (n=5, *P=0.002). A 2.1-fold increase in G3923 methylation is observed after scaRNA2 knockdown compared to control 2 (n=4, *P=0.002).