Rocaglamide a

HeLa, 293A, 293T, Huh7, BHK-21, and Vero cells were cultured in Dulbecco’s modified Eagle’s medium supplemented with 10% fetal calf serum, 100 U/ml penicillin, and 100 μg/ml streptomycin at 5% CO₂ and 37 °C. JEV AT31 and DENV 2 NGC strains (ATCC VR-1584) were cultured in 293A and BHK-21 cells, respectively. DBeQ (Sigma), Eer1 (Sigma), MDBN (StressMarq Biosciences), and rocaglamide A (Sigma) were dissolved in dimethyl sulfoxide. The culture medium of mock-treated cells contained the highest inhibitor concentration used in the respective assay. To induce the formation of SGs, the cells were cultured in a growth medium containing 100 μM sodium arsenite. The siRNAs, plasmids, and antibodies are listed in the Supplemental Information (Tables S1–S3). Virus titration was performed with a focus forming assay as described (46 (link)). For the infection study, viruses were inoculated at a multiplicity of infection of 0.3 for propagation and 1.0 for imaging experiments, including the quantification of SGs.

Sterols, 7-α-aminocholesterol, cholesterol, 6-ketocholestanol and rocaglamide A were purchased from Sigma-Aldrich (St. Louis, MO, USA) under the references R207578, C3045, K1250 and SML0656, respectively. The structures of the compounds used are shown in Figure 1. The compounds were dissolved in dimethyl sulfoxide (DMSO; Sigma-Aldrich, St. Louis, MO, USA) to make stock solutions.