ExtData_Figure_3_replicates

(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 STOML2 or YKT6 ~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. R# --> replicate number; LG### --> experiment number. .sgd files are the original source files acquired with the Syngene Imager.