Fig. 3.
Effect of mRNA cap methylation on PCIF1 modification of VSV mRNA. (A) The indicated 293T cells were infected with VSV-LG1670A at a MOI of 3, and viral RNA was radiolabeled, extracted, and analyzed by 2D-TLC as in Fig. 2A (representative images, n = 3). (B) mRNA synthesized in vitro by VSV-LG1670A mRNA was incubated with purified PCIF1 and analyzed as in Fig. 2C (representative image, n = 3). (C) The indicated 293T cells were infected with VSV-LG4A as in A (representative images, n = 3). (D) mRNA synthesized by VSV in vitro in the presence of 200 µM S-adenosyl-homocysteine was used as substrate for PCIF1 in vitro and analyzed as in B (representative image, n = 3).