Figure 3.

The interaction between RBM24 and HCV RNA sequences. (A and B) Huh7.5.1 cells were transfected with pFlag-RBM24 and infected with Jc1 at an MOI of 0.1. The cells were harvested at 72 hpi as described in experimental procedures and immunoprecipitated with either an anti-Flag mouse monoclonal antibody (Flag) or a nonspecific mouse control antibody (IgG). Precipitated HCV RNA was detected by qRT-PCR (A) or RPA (B), and the precipitated NS3, RBM24 and actin were detected by Western blot (B). (C) Huh7.5.1 cells were transfected with vectore or pHA-RBM24 and then infected with Jc1 at an MOI of 1. The localization of double strand RNA and RBM24 protein were detected by immunofluorescence using the J2 antibody and HA antibody respectively. The nuclei were stained with Hoechst 33258. (D–F) A lysate of 293T cells transfected with pFlag-RBM24 was crosslinked with the indicated ³²P-labeled HCV fragments. The crosslinked nucleotides-proteins were immunoprecipitated with either an anti-Flag mouse monoclonal antibody (Flag) or a nonspecific mouse control antibody (IgG) and detected by autoradiography. The input proteins were detected by Western blot. (G) A lysate of 293T cells transfected with pcDNA3.1-RBM24 was incubated with the indicated biotin-labeled HCV fragments and affinity-precipitated with streptavidin beads. RBM24 protein was detected by Western blot