Figure 3

Effect of hnRNP A1 on the synthesis of G and SG RNA in vitro. (A) Western blot to examine the expression of nsP4. BSC40 cells were transfected with no siRNA or siRNA targeting hnRNP A1 for 3 days as described under Materials and Methods. Cells were co-infected with recombinant vaccinia viruses expressing T7 RNA polymerase, SV nsP123 and SV nsP4 at an moi of one for 24 h. Cell lysates were prepared and subjected to 12% SDS-PAGE electrophoresis. Western blot was carried out using antibody to nsP4. Expression of actin was examined by Western blot as a control. (B) in vitro G and SG RNA synthesis. hnRNP A1 was depleted from BSC40 cells as described above. Cells were co-infected with recombinant vaccinia viruses expressing T7 RNA polymerase, SV nsP123 and SV nsP4 and P15 fractions were prepared as described under Materials and Methods. P/T were prepared and standard reaction conditions were used as described under Materials and Methods.