ExtData_Figure_2_images_for_paper

<em>(A) Xrn1 knock out A549 cells were infected with WT PR8 or PR8-PA(∆X), or mock infected. RNA was isolated, 5’ RACE was performed using primers specific for </em>BCAP31<em>, </em>TUBA1B<em>, </em>INSIG1<em> or </em>SLC7A5<em>, and the PCR products were run on an agarose gel. Primers were positioned ~200-300 nucleotides downstream of the predicted cut sites. </em> <em>(C) HEK293T ishXrn1 cells were treated with no drug or doxycycline for 3-4 days to induce shRNAs against Xrn1, then protein lysates were collected and probed with antibodies against Xrn1, or b-tubulin as a loading control to check for efficient knock down of Xrn1. Images are representative of 3 experiments. </em> <em>(E) HEK293T ishXrn1 cells were treated with doxycycline for 3-4 days to induce knock down of Xrn1, then transfected with luciferase reporters containing 99 bp insertions from the indicated genes (</em>INSIG1, SLC7A5, STOML2, YKT6, TUBA1B, and BCAP31)<em>, and where indicated, with PR8 PA-X. RNA was extracted and used to run 5’ RACE. For all gels, the DNA bands were purified and sequenced to confirm their identities. Images are representative of 3 experiments or 2 experiments for luciferase + </em>INSIG1/SLC7A5<em>. The SLC7A5 sequence was only consistently cut when inside the luciferase reporter.</em> <em>Additional replicates can be found in ExtData_Figure_2_replicates dataset.</em> <em>R# --&gt; replicate number; LG### --&gt; experiment number.</em> <em>.sgd files are the original source files acquired with the Syngene Imager. </em>