(A) Northerns for β-GAP and control reporter mRNAs in inputs (0.5%) or coprecipitated with Flag-Upf1. Schematic of the β-GAP mRNAs used in RNA-IPs is shown below Northerns. HP denotes a stable RNA hairpin in the 5′ UTR that blocks translation. Graphs on the right represent mean IP recovery over input of β-GAP mRNA normalized by recovery of the internal control after subtraction of background from negative control IPs, +/− SEM for triplicate biological repeats.
(B) Similar to panel A for β-globin RNA-IPs, except performed in the presence of Smg6 depletion to prevent Smg6-mediated cleavage of the β-globin PTC mRNA (see Figure S1).
Asterisks denote P-values: *≤0.1, **≤0.05, *** ≤0.01 (paired student’s t-test, two-tailed).
See also Figure S4