Detection and quantification of glycosylated queuosine modified tRNAs by acid denaturing and APB gels

Q and glycosylated Q-modification kinetics of HEK293T cells by APB and acid denaturing gels. (A) Time course of individual Q or glycosylated Q-modified tRNA. At t = 0, 1 µM queuine was added to the medium of 0Q cells, total RNA was extracted after designated time points in hours. tRNA^His and tRNA^Asn are from APB gels, tRNA^Tyr and tRNA^Asp are from acid denaturing gels. G: unmodified tRNA, Q: Q-modified tRNA, mQ: mannosyl-Q, gQ: galactosyl-Q. (B) Quantitative modification fraction over time for individual tRNAs, from 1 µM queuine data. For tRNA^His and tRNA^Asn, the modification fraction represents Q; for tRNA^Asp and tRNA^Tyr, the modification fraction represents the sum of the Q and glycosylated Q modifications. (C,D) Same as A,B except at t = 0, 10 nM queuine was added to the medium of 0Q cells.