Both SNORD97 and SCARNA97 participate in 2′-O-methylation of tRNAMet(CAT). (A) Northern blot analysis of SNORD97 and SCARNA97 expression in HAP1, SNORD97 knockout (SNO97-KO), SCARNA97 knockout (SCA97-KO), and SNORD97 + SCARNA97 knockout (SNO97 + SCA97-KO) cells. The 7SK and U1 snRNAs are positive controls. (B) Two-dimensional structure of human elongator tRNAMet(CAT) with modified nucleotides (Boccaletto et al. 2018). Position of the oligonucleotide used for primer extension is shown. (C) Mapping of 2′-O-methylation of tRNAMet(CAT). Cellular RNAs extracted from the indicated cell lines were subjected to partial alkaline hydrolysis. Terminally labeled oligonucleotide primer was annealed to each RNA sample and extended with reverse transcriptase. (Lanes U,G,C,A) Dideoxy sequencing ladders.