Effect of overexpression of DRBD18 methyl mutants on RNA editing. (A) Cartoon depicting DRBD18 domains and sites of methylation and mutations. (RRM) RNA recognition motif. Numbers indicate amino acid residues. Stars indicate sites of methylated arginines. (B, top) Western blot showing degree of 2XMyc-DRBD18 overexpression in three biological replicates of uninduced (−Doxy) and induced (+Doxy) DRBD18(R→K) cells. Levels of total DRBD18 are shown on the right. (Bottom) qRT-PCR analysis of RNA isolated from uninduced (−Doxy) and induced (+Doxy) DRBD18(R→K) overexpression cells using primer sets designed to detect total, preedited, and edited mRNA of a subset of mitochondrial transcripts. The relative abundance of each transcript in induced versus uninduced RNAi cells is shown, and the RNA levels were normalized to 18S rRNA. Three biological replicates, each with three technical replicates, were performed. (C) As in B using DRBD18(R→F) cells.
