Figure 5A mRNA of IFNAR1, but not IFNAR2 or STAT1, was stabilized by RBM47 ectopic expression in 293T cells with or without IFN-α stimulation. Cells were treated with IFN-α or PBS for 6 h, and the relative gene expression levels were determined using qRT-PCR.The PCR results are represented as the means ± SD of n = 3 biological replicates. Data information: The data shown are representative of n = 3 independent experiments. NS, non-significant; **P ≤ 0.01, and ***P ≤ 0.001 (Student's t-test). The data shown are representative of three independent experiments. B, mRNA levels of IFNAR1 were analyzed in WT and RBM47 knockout 293T cells with or without IFN-α stimulation. The relative density of IFNAR1 protein in the control group was set to "1." The PCR results are represented as the means ± SD of n = 3 biological replicates. Data information: The data shown are representative of n = 3 independent experiments. NS, non-significant; **P ≤ 0.01, and ***P ≤ 0.001 (Student's t-test). The data shown are representative of three independent experiments. , C protein levels of IFNAR1 were analyzed in WT and RBM47 knockout 293T cells with or without IFN-α stimulation. The relative density of IFNAR1 protein in the control group was set to "1." The PCR results are represented as the means ± SD of n = 3 biological replicates. Data information: The data shown are representative of n = 3 independent experiments. NS, non-significant; **P ≤ 0.01, and ***P ≤ 0.001 (Student's t-test). The data shown are representative of three independent experiments. D IFNAR1 mRNA expression in PMs from WT and RBM47+/- mice treated with or without IFN-α. The PCR results are represented as the means ± SD of n = 3 biological replicates. Data information: The data shown are representative of n = 3 independent experiments. NS, non-significant; **P ≤ 0.01, and ***P ≤ 0.001 (Student's t-test). The data shown are representative of three independent experiments. E Flow cytometry analysis of IFNAR1 in the cytoplasmic membrane of BMDMs from WT and RBM47+/- mice. The cells was treated with PBS or IFN-α for 6 h. Data are represented as the means ± SD of n = 3 biological replicates. Data information: The data shown are representative of n = 3 independent experiments. NS, non-significant; **P ≤ 0.01, and ***P ≤ 0.001 (Student's t-test). The data shown are representative of three independent experiments. F, G Protein levels of IFNAR1 and phosphorylation levels of TYK2, STAT1, and STAT2 in RBM47-overexpressing (F) or knockdown (G) 293T cells. Cells were treated with PBS or IFN-α for 6 h, and the relative band densities were analyzed using ImageJ software and normalized to GAPDH. Data information: The data shown are representative of n = 3 independent experiments. NS, non-significant; **P ≤ 0.01, and ***P ≤ 0.001 (Student's t-test). The data shown are representative of three independent experiments. H ISRE luciferase was tested to investigate the effects of RBM47 or its variants on ISG activation stimulated by IFN-α.Data are represented as the means ± SD of n = 3 biological replicates. Data information: The data shown are representative of n = 3 independent experiments. NS, non-significant; **P ≤ 0.01, and ***P ≤ 0.001 (Student's t-test). The data shown are representative of three independent experiments. I, J Effect of overexpression of WT or RBM47 mutants on VSV replication. 293T cells were transfected with RBM47-Flag or two mutants for 24 h and then infected with GFP-expressing VSV (MOI = 0.1) for another 12 h. The replication of VSVG was observed using fluorescence microscopy via GFP signal (I)(Scale bar, 50 μm), and protein levels were determined using western blot (J).The relative band densities were analyzed using ImageJ software and normalized to GAPDH. The relative density of VSVG protein in the control group was set to "1." Data information: The data shown are representative of n = 3 independent experiments. NS, non-significant; **P ≤ 0.01, and ***P ≤ 0.001 (Student's t-test). The data shown are representative of three independent experiments.