FLAG-RIG-I and FLAG-GFP, cloned into the pcDNA5 FRT/TO vector, have been previously described10 (link). pCMV6-Entry plasmids encoding Myc-DDK-tagged human RPL5, MRPL18, and TST were purchased from OriGene. The DNA construct encoding U6 promoter-expressed RNA5SP141 was generated by PCR using the following forward (F) and reverse (R) primers: F1 primer: 5′-TGGAAAGGACGAAACACCGTCTACGGCCATACCACCCTGAACGCGCCCGATCTCGTC-3′; F2 primer: 5′-CGCGCCCGATCTCGTCTGATCTCGGAAGCTAAGCAGGGTTGGGCCTGGTTAGTACTTGGATGGGAGAAATACATCCAAAAACACGATGACTCACATG-3′; R primer: 5′-CATGTGAGTCATCGTGTTTTTGGATG-3′. The DNA construct encoding U6 promoter-expressed RABVINT was generated using the following F and R primers: F1 primer: 5′-TGGAAAGGACGAAACACCGGGCCAGGATGGCGGCTCAAACTGCTTCTGGCCCTCCAGCCCTTGAATGGTCGGCCACC-3′; R primer: 5′-GGTGGCCGACCATTCAAGGGCT-3′.
Transient transfection of cells was performed using calcium phosphate (Clontech), linear polyethylenimine (PEI (Polysciences, Inc.) at 1 mg/ml in 10 mM Tris pH 6.8), Lipofectamine and Plus reagent or Lipofectamine 2000 (both Life Technologies), according to the manufacturer’s instructions.
Transient transfection of cells was performed using calcium phosphate (Clontech), linear polyethylenimine (PEI (Polysciences, Inc.) at 1 mg/ml in 10 mM Tris pH 6.8), Lipofectamine and Plus reagent or Lipofectamine 2000 (both Life Technologies), according to the manufacturer’s instructions.