Q-modification kinetics of tRNAAsp and tRNATyr in HEK293T cells by APB gels. (A) At t = 0, 1 µM queuine was added to the medium of 0Q cells, total RNA was extracted after designated time points in hours. G: unmodified tRNA, Q: Q-modified tRNA, mQ: mannosyl-Q, gQ: galactosyl-Q. The reversal to fast migration in later time points corresponds to gQ or mQ products as indicated by the acid denaturing gel results from Figure 3. (B) Quantitative modification fraction over time for individual tRNAs, from 1 µM queuine data. For comparison the total modification fraction from acid denaturing gel results in Figure 3 are also shown. (C,D) Same as A,B except at t = 0, 10 nM queuine was added to the medium of 0Q cells.
