Figure 5.

The ARO4pG mRNA is destabilized after rapamycin treatment by having a prematurely short poly(A) tail. Transcriptional pulse-chase analysis of the ARO4pG mRNA after treatment with either vehicle (− rapamycin) or rapamycin (+ rapamycin) for 60 min is shown. In each sample, the ARO4pG mRNA was cleaved with oCD163 and RNAse H to resolve the length of its poly(A) tail. The poly(A) tail was removed by treatment with oCD163, oligo dT, and RNAse H (dT lane). Full-length and fragment species were detected with oRP127 and are indicated at the right of the figure. Poly(A) tail length is marked by A50 and A0 indicators. The marker (M) lane contains 5′ end-labeled HinfI ϕχ174 DNA, and sizes are indicated to the left of the figure.