TRIM2 decreases JUNV entry into cells.

(A) TRIM2 knockdown does not affect virus binding to cells. U2OS cells were transfected with a TRIM2 siRNA and incubated with FITC-labeled Candid 1. Shown is a representative FACS plot. This experiment was performed twice with similar results. (B) Cells overexpressing hTRIM2 or mTRIM2 or GFP (Control) were incubated with Candid 1, and after a 1-hr incubation at 37°C, virus was stripped from cells, and RNA was isolated and analyzed for viral RNA by RT-qPCR. Shown are the averages ± SD of 6 independent experiments. P values were determined by unpaired t tests; ***P ≤ 0.0002; ****P ≤ 0.0001. See also S4 Fig. (C) The same experiment was performed with primary bone marrow–derived macrophages isolated from mice of the indicated genotype. Values represent the mean ± SD in 2 independent experiments with triplicate experimental replicates. FACS, fluorescence-activated cell sorting; FITC, fluorescein isothiocyanate; GFP, green fluorescent protein; hTRIM2, human TRIM2; JUNV, Junín virus; mTRIM2, mouse TRIM2; RT-qPCR, real-time quantitative PCR; siRNA, small interfering RNA; TfR1, transferrin receptor 1; WT, wild type.