cometadata/datacite-titles-descriptions-related-identifiers huggingface dataset:
DOI: 10.6084/m9.figshare.22714843
title: ExtData_Figure_5_replicates

description: <em>(A) HEK293T ishXrn1cells were treated with doxycycline for 3-4 days to induce knock down of Xrn1, then transfected with luciferase reporters containing insertions of the indicated lengths from the BCAP31 and SLC7A5 genes, with or without PR8 PA-X. RNA was extracted and used to run 5’ RACE. (B) HEK293T ishXrn1cells were treated with doxycycline for 3-4 days to induce knock down of Xrn1, then transfected with luciferase reporters containing 15 bp insertions from the </em>STOML2<em> gene with or without mutations to disrupt/restore the hairpin structure. 24 hours post transfection, cells were infected with WT PR8 or PR8-PA(∆X), or mock infected overnight. RNA was then extracted and used to run 5’ RACE. For all gels, the DNA bands were purified and sequenced to confirm their identities.</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>
DOI: 10.6084/m9.figshare.22714843.v1
title: ExtData_Figure_5_replicates

description: <em>(A) HEK293T ishXrn1cells were treated with doxycycline for 3-4 days to induce knock down of Xrn1, then transfected with luciferase reporters containing insertions of the indicated lengths from the BCAP31 and SLC7A5 genes, with or without PR8 PA-X. RNA was extracted and used to run 5’ RACE. (B) HEK293T ishXrn1cells were treated with doxycycline for 3-4 days to induce knock down of Xrn1, then transfected with luciferase reporters containing 15 bp insertions from the </em>STOML2<em> gene with or without mutations to disrupt/restore the hairpin structure. 24 hours post transfection, cells were infected with WT PR8 or PR8-PA(∆X), or mock infected overnight. RNA was then extracted and used to run 5’ RACE. For all gels, the DNA bands were purified and sequenced to confirm their identities.</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>
DOI: 10.6084/m9.figshare.22714849
title: ExtData_Figure_6_images_for_paper

description: <em>(D-F) Xrn1 knock out A549 cells were infected with WT PR8 or PR8-PA(∆X), or mock infected. 5’RACE was performed and PCR products were run on an agarose gel. </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>
DOI: 10.6084/m9.figshare.22714804.v1
title: ExtData_Figure_4_images_for_paper

description: <em>(F) HEK293T ishXrn1cells 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 BCAP31 or TUBA1B, with or without PR8 PA-X. RNA was extracted and used to run 5’ RACE. DNA bands were purified and sequenced to confirm their identities, and images are representative of 3 experiments.</em> <em>Additional replicates can be found in ExtData_Figure_4_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>
DOI: 10.6084/m9.figshare.22714813
title: ExtData_Figure_4_replicates

description: <em>(F) HEK293T ishXrn1cells 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 BCAP31 or TUBA1B, with or without PR8 PA-X. RNA was extracted and used to run 5’ RACE. DNA bands were purified and sequenced to confirm their identities.</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>
DOI: 10.6084/m9.figshare.22714822.v1
title: ExtData_Figure_4_qPCR

description: <em>(E) HEK293T ishXrn1cells 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 BCAP31 or TUBA1B, with or without PR8 PA-X. </em> <em>RNA was extracted and luciferase mRNA levels were quantified by qRT-PCR and normalized to 18S. </em> <em>R# --&gt; replicate number; LG### --&gt; experiment number.</em> <br>
DOI: 10.6084/m9.figshare.22714828.v1
title: ExtData_Figure_5_images_for_paper

description: <em>(A) HEK293T ishXrn1cells were treated with doxycycline for 3-4 days to induce knock down of Xrn1, then transfected with luciferase reporters containing insertions of the indicated lengths from the BCAP31 and SLC7A5 genes, with or without PR8 PA-X. RNA was extracted and used to run 5’ RACE. (B) HEK293T ishXrn1cells were treated with doxycycline for 3-4 days to induce knock down of Xrn1, then transfected with luciferase reporters containing 15 bp insertions from the </em>STOML2<em> gene with or without mutations to disrupt/restore the hairpin structure. 24 hours post transfection, cells were infected with WT PR8 or PR8-PA(∆X), or mock infected overnight. RNA was then extracted and used to run 5’ RACE. For all gels, the DNA bands were purified and sequenced to confirm their identities, and images are representative of 3 experiments (B) or 2 experiments (A). </em> <em>Additional replicates can be found in ExtData_Figure_5_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>
DOI: 10.6084/m9.figshare.22714849.v1
title: ExtData_Figure_6_images_for_paper

description: <em>(D-F) Xrn1 knock out A549 cells were infected with WT PR8 or PR8-PA(∆X), or mock infected. 5’RACE was performed and PCR products were run on an agarose gel. </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>
DOI: 10.6084/m9.figshare.22714804
title: ExtData_Figure_4_images_for_paper

description: <em>(F) HEK293T ishXrn1cells 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 BCAP31 or TUBA1B, with or without PR8 PA-X. RNA was extracted and used to run 5’ RACE. DNA bands were purified and sequenced to confirm their identities, and images are representative of 3 experiments.</em> <em>Additional replicates can be found in ExtData_Figure_4_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>
DOI: 10.6084/m9.figshare.22714813.v1
title: ExtData_Figure_4_replicates

description: <em>(F) HEK293T ishXrn1cells 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 BCAP31 or TUBA1B, with or without PR8 PA-X. RNA was extracted and used to run 5’ RACE. DNA bands were purified and sequenced to confirm their identities.</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>
DOI: 10.6084/m9.figshare.22714636
title: ExtData_Figure_3_replicates

description: <em>(A-C) 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, and where indicated, with WT PA-X from the PR8 strain (A-C), the catalytic mutant PR8 PA-X D108A (A), PR8 PA with a mutation to reduce frameshifting and prevent PA-X production (PA(fs)) (A), herpes simplex virus 1 (HSV-1) vhs or Kaposi’s sarcoma-associated herpes virus (KSHV) SOX (B), or WT PA-X from the Perth influenza strain (C). RNA was extracted and used to run 5’ RACE. (D) Xrn1 knock out A549 cells were infected with WT PR8, Perth or pH1N1pdm09 or the corresponding PA(∆X) mutants, or mock infected. 5’ RACE was then performed using primers specific for </em>STOML2<em> or </em>YKT6<em> ~250-300 nt downstream of the predicted cut sites. The PCR products were run on an agarose gel. For all gels, the DNA bands were purified and sequenced to confirm their identities. </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>
DOI: 10.6084/m9.figshare.22714570
title: ExtData_Figure_2_replicates

description: <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.</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. The SLC7A5 sequence was only consistently cut when inside the luciferase reporter.</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>
DOI: 10.6084/m9.figshare.22714570.v1
title: ExtData_Figure_2_replicates

description: <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.</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. The SLC7A5 sequence was only consistently cut when inside the luciferase reporter.</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>
DOI: 10.6084/m9.figshare.22714582
title: ExtData_Figure_7_images_for_paper

description: <em>(C-D) HEK293T ishXrn1 cells were treated with doxycycline for 3-4 days to induce knock down of Xrn1, then transfected with luciferase reporters containing the 99 bp insertions in </em>STOML2 or YKT6<em> either in an exon or an intron, with or without PR8 PA-X. (B) RNA was extracted and </em> <em>used to run 5’ RACE. PCR products were separated on an agarose gel. Gel images are representative of 3 experiments.</em> <em>Additional replicates can be found in ExtData_Figure_7_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>
DOI: 10.6084/m9.figshare.22714588
title: ExtData_Figure_7_replicates

description: (C-D) <em>HEK293T ishXrn1 cells were treated with doxycycline for 3-4 days to induce knock down of Xrn1, then transfected with luciferase reporters containing the 99 bp insertions in the indicated genes either in an exon or an intron, with or without PR8 PA-X. RNA was extracted and</em> <em>used to run 5’ RACE. PCR products were separated on an agarose gel. </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>
DOI: 10.6084/m9.figshare.22714588.v1
title: ExtData_Figure_7_replicates

description: (C-D) <em>HEK293T ishXrn1 cells were treated with doxycycline for 3-4 days to induce knock down of Xrn1, then transfected with luciferase reporters containing the 99 bp insertions in the indicated genes either in an exon or an intron, with or without PR8 PA-X. RNA was extracted and</em> <em>used to run 5’ RACE. PCR products were separated on an agarose gel. </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>
DOI: 10.6084/m9.figshare.22714609
title: ExtData_Figure_3_images_for_paper

description: <em>(A-C) 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, and where indicated, with WT PA-X from the PR8 strain (A-C), the catalytic mutant PR8 PA-X D108A (A), PR8 PA with a mutation to reduce frameshifting and prevent PA-X production (PA(fs)) (A), herpes simplex virus 1 (HSV-1) vhs or Kaposi’s sarcoma-associated herpes virus (KSHV) SOX (B), or WT PA-X from the Perth influenza strain (C). RNA was extracted and used to run 5’ RACE. (D) Xrn1 knock out A549 cells were infected with WT PR8, Perth or pH1N1pdm09 or the corresponding PA(∆X) mutants, or mock infected. 5’ RACE was then performed using primers specific for </em>STOML2<em> or </em>YKT6<em> ~250-300 nt downstream of the predicted cut sites. The PCR products were run on an agarose gel. For all gels, the DNA bands were purified and sequenced to confirm their identities, and images are representative of 3 experiments. </em> <em>Additional replicates can be found in ExtData_Figure_3_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>
DOI: 10.6084/m9.figshare.22714609.v1
title: ExtData_Figure_3_images_for_paper

description: <em>(A-C) 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, and where indicated, with WT PA-X from the PR8 strain (A-C), the catalytic mutant PR8 PA-X D108A (A), PR8 PA with a mutation to reduce frameshifting and prevent PA-X production (PA(fs)) (A), herpes simplex virus 1 (HSV-1) vhs or Kaposi’s sarcoma-associated herpes virus (KSHV) SOX (B), or WT PA-X from the Perth influenza strain (C). RNA was extracted and used to run 5’ RACE. (D) Xrn1 knock out A549 cells were infected with WT PR8, Perth or pH1N1pdm09 or the corresponding PA(∆X) mutants, or mock infected. 5’ RACE was then performed using primers specific for </em>STOML2<em> or </em>YKT6<em> ~250-300 nt downstream of the predicted cut sites. The PCR products were run on an agarose gel. For all gels, the DNA bands were purified and sequenced to confirm their identities, and images are representative of 3 experiments. </em> <em>Additional replicates can be found in ExtData_Figure_3_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>
DOI: 10.6084/m9.figshare.22710316.v1
title: ExtData_Figure_1_replicates

description: <em>(B) Protein lysates of Xrn1 knock out A549 cells infected with WT PR8 or PR8-PA(∆X), or mock infected, were probed with antibodies against PR8 PA or b-tubulin as a loading control. Images are representative of 2 experiments. </em> <em>(C) Protein lysates of WT or Xrn1 knock out (ko) A549 cells were probed with antibodies against Xrn1 or b-tubulin as a loading control to check for loss of Xrn1.</em> <em>R# --&gt; replicate number; LG### --&gt; experiment number.</em> <em>.sgd files are original source files acquired with Syngene Imager.</em>
DOI: 10.6084/m9.figshare.22714567
title: ExtData_Figure_2_images_for_paper

description: <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>
DOI: 10.6084/m9.figshare.22714579
title: ExtData_Figure_7_qPCR

description: <em>(B) Testing of luciferase reporters with sequence insertions in the introns, as shown in diagram in A. HEK293T ishXrn1 cells were treated with doxycycline for 3-4 days to induce knock down of Xrn1, then transfected with luciferase reporters containing the 99 bp insertions in </em>STOML2 or YKT6 <em>either in an exon or an intron, with or without PR8 PA-X. RNA was extracted and luciferase mRNA levels were quantified by qRT-PCR and normalized to 18S. Two sets of luciferase primers were used - either both annealing to exons or one to exon and one to intron, to measure levels of the processed mRNA and unspliced pre-mRNA, respectively.</em>
DOI: 10.6084/m9.figshare.22710250.v1
title: ExtData_Figure_1_images_for_paper

description: <em>B) Protein lysates of Xrn1 knock out A549 cells infected with WT PR8 or PR8-PA(∆X), or mock infected, were probed with antibodies against PR8 PA or b-tubulin as a loading control. Images are representative of 2 experiments. </em> <em>(C) Protein lysates of WT or Xrn1 knock out (ko) A549 cells were probed with antibodies against Xrn1 or b-tubulin as a loading control to check for loss of Xrn1. Images are representative of 2 experiments. </em> <em>R# --&gt; replicate number; LG### --&gt; experiment number. Additional replicates are found in ExtData_Figure_1_replicates dataset.</em> <em>.sgd files are original source files acquired with Syngene Imager.</em>
DOI: 10.6084/m9.figshare.22710316
title: ExtData_Figure_1_replicates

description: <em>(B) Protein lysates of Xrn1 knock out A549 cells infected with WT PR8 or PR8-PA(∆X), or mock infected, were probed with antibodies against PR8 PA or b-tubulin as a loading control. Images are representative of 2 experiments. </em> <em>(C) Protein lysates of WT or Xrn1 knock out (ko) A549 cells were probed with antibodies against Xrn1 or b-tubulin as a loading control to check for loss of Xrn1.</em> <em>R# --&gt; replicate number; LG### --&gt; experiment number.</em> <em>.sgd files are original source files acquired with Syngene Imager.</em>
DOI: 10.6084/m9.figshare.22710586
title: ExtData_Figure_9_TCID50images

description: <em>(C) MDCK cells were infected at MOI 0.05 with the following viruses: </em> <em>WT PR8 and PR8-PA(∆X) strains with no insert, PR8 with WT or mutant </em>STOML2<em> sequences inserted in the WT PR8 or PR8-PA(∆X) background. Mutant </em>STOML2<em> sequences contain the GCUG to UAGC mutation preventing PA-X cleavage. Supernatants were collected at 1, 8, 24 and 48 hours post infection and viral titers were quantified by TCID50.</em>
DOI: 10.6084/m9.figshare.22710586.v1
title: ExtData_Figure_9_TCID50images

description: <em>(C) MDCK cells were infected at MOI 0.05 with the following viruses: </em> <em>WT PR8 and PR8-PA(∆X) strains with no insert, PR8 with WT or mutant </em>STOML2<em> sequences inserted in the WT PR8 or PR8-PA(∆X) background. Mutant </em>STOML2<em> sequences contain the GCUG to UAGC mutation preventing PA-X cleavage. Supernatants were collected at 1, 8, 24 and 48 hours post infection and viral titers were quantified by TCID50.</em>
DOI: 10.6084/m9.figshare.22714567.v1
title: ExtData_Figure_2_images_for_paper

description: <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>
DOI: 10.6084/m9.figshare.22710208
title: ExtData_Figure_2_qPCR

description: <em>(B) Xrn1 knock out A549 cells were infected with WT PR8 or PR8-PA(∆X), or mock infected.</em> <em>PR8 HA RNA levels were quantified by qRT-PCR and normalized to 18S.</em> <em>(D) 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 </em> INSIG1,SLC7A5,STOML2,YKT6,TUBA1B or BCAP31<em> and where indicated, with PR8 PA-X. RNA was extracted and luciferase mRNA levels were quantified by qRT-PCR and normalized to 18S</em> . <em>R# --&gt; replicate number; LG### --&gt; experiment number</em>
DOI: 10.6084/m9.figshare.22710208.v1
title: ExtData_Figure_2_qPCR

description: <em>(B) Xrn1 knock out A549 cells were infected with WT PR8 or PR8-PA(∆X), or mock infected.</em> <em>PR8 HA RNA levels were quantified by qRT-PCR and normalized to 18S.</em> <em>(D) 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 </em> INSIG1,SLC7A5,STOML2,YKT6,TUBA1B or BCAP31<em> and where indicated, with PR8 PA-X. RNA was extracted and luciferase mRNA levels were quantified by qRT-PCR and normalized to 18S</em> . <em>R# --&gt; replicate number; LG### --&gt; experiment number</em>