Fig. 1
From: Modification of messenger RNA by 2′-O-methylation regulates gene expression in vivo
2′-O-methylation on Pxdn mRNA requires fibrillarin. a 2′-O-methylation (Nm) can occur in combination with any base. Nucleoside Am is shown, compared to base-modified m6A. b UPLC-MS/MS quantification of modified nucleosides on total RNA, mRNA after two rounds of oligo-dT selection, and mRNA further purified by rRNA depletion. c Predicted interaction between human snoRNA U32A (U32A) and peroxidasin (Pxdn). A3150 is the predicted Nm target (red), which is the first position of a lysine codon (K). d Illustration of RTL-P method. The amount of qPCR product from the low dNTP reactions of two different samples can be compared to establish the relative “RTL-P efficiency (RQ)”. RQ = relative quantity. See Methods for detail. e–f Fbl knockdown (KD) in HeLa cells, vs. negative control (ctrl) using siRNA for 48 h. n = 3 independent experiments, SEM error bars, *p < 0.05 by unpaired t-test. e Fbl KD leads to loss of Nm on Pxdn mRNA (p = 0.0005). f Fbl KD reduces Pxdn mRNA levels, normalized to Rplp0 control transcript (p = 0.0002)
