We followed previously described methods (57). Briefly, MXA reporter adults were outcrossed with wt AB zebrafish and eggs were injected at the one-cell stage with 1 nl of a 0.5 mM solution of MO (Gene Tools), targeting a trim8a or trim8b splice site (fig. S2) or a control MO with no known target. Eggs were then bleached and incubated in Volvic water at 24° or 28°C, according to the desired speed of development. One day later, 0.003% 1-phenyl-2-thiourea (Sigma-Aldrich) was added to prevent pigmentation; this was kept for all further steps. When the larvae had reached the 3-dpf stage, they were dechorionated if needed, anesthetized with tricaine (Sigma-Aldrich, A-5040), and inoculated intravenously by targeting the large vessels caudal to the cloaca, with ~200 PFU of CHIKV-GFP, as described in (32). After a rinse, these larvae were maintained at 28°C in individual wells of a 24-well plate and inspected at least daily with a stereomicroscope. Quantitative assessment of the clinical status, performed 3 days after inoculation, was based on a precise list of criteria, as previously described (32). Briefly, clinical signs were assessed blindly, yielding a disease score ranging from 0 (no disease sign) to 15 (dead or terminally ill). The signs evaluated included the following: ability to maintain equilibrium, response to touch, body shape, blood flow, cardiac rhythm, presence of edema, inflation of the swim bladder, and opacity of the yolk.

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