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To deal with this matter, we titrated the amount of cycloheximide so you can a sandwich-lethal concentration (0

To deal with this matter, we titrated the amount of cycloheximide so you can a sandwich-lethal concentration (0

Contour step three-supply studies dos

Origin investigation getting Figure 3B, C, D–Grams, I and you can J: decay kinetics regarding selected mRNAs within the structure given translational inhibitors.

Profile step three-source analysis step three

To begin to test these predictions, we directly perturbed the process of translation elongation and observed the effects on mRNA decay. Consistent with previous reports, we found that treating cells with a strong dose (50 ?g/mL) of the elongation inhibitor cycloheximide resulted in stabilized CIS3 and RPL25 transcripts and had no significant effect on ACT1 mRNA stability (Figure 3-figure supplement 1A) (Beelman and Parker, 1994). A potentially problematic aspect of these experiments is that high doses of cycloheximide completely halt translation thus shutting down a myriad of cellular processes, which in turn could lead to indirect effects. 2 ?g/mL) and assayed the effect of this level of elongation inhibition on transcript stability (Figure 3-figure supplement 2H).