FIGURE 2.
Optimization of 4sU-labeling of mRNA in cells. (A) Effect of increasing 4sU concentrations on cell viability. HEK-293T cells were grown 24 h in the indicated concentrations of 4sU. Viability was measured using CellTiter-Glo. Error bars are SD, n = 3. (B) Concentrations of 4sU required to impede reverse transcription of endogenous mRNAs. RNA was isolated from HEK-293T cells treated with the indicated concentrations of 4sU for 4 h, and then treated with NEM and reverse transcribed. Levels of the indicated mRNAs were monitored by qPCR analysis of cDNA. Error bars are SD, n = 3.
