Figure 2: RIPK signaling exerts CNS-intrinsic restriction of ZIKV replication.

(A-C) Survival analysis in Ripk3^−/− (A), Ripk1^KD/KD (B), or Mlkl^−/− (C) mice and congenic controls following intracranial inoculation with ZIKV-MR766. N= 10–16 mice/genotype.

(D) Survival analysis in Ripk3-2xFV^fl/fl Mox2-Cre⁺ and Ripk3-2xFV^fl/fl Mox2-Cre⁺ Mlkl^−/− mice along with Cre⁻ littermate controls following intracranial inoculation with ZIKV-MR766. N= 9–15 mice/genotype.

(E-G) Ripk3^−/− (E), Ripk1^KD/KD (F), or Mlkl^−/− (G) mice and congenic controls were infected intracranially with ZIKV-MR766. On indicated days following infection, whole brains were assayed for ZIKV titers via plaque assay.

(H) Analysis of brain viral burden in Ripk3-2xFV^fl/fl Mox2-Cre⁺ and Ripk3-2xFV^fl/fl Mox2-Cre⁺ Mlkl^−/− mice along with Cre⁻ littermate controls following intracranial ZIKV-MR766 infection, as in (E-G).

(I-K) Multistep viral growth curve analysis (MOI 0.1) in primary cortical neurons isolated from Ripk3^−/− (I), Ripk1^KD/KD (J), or Mlkl^−/− (K) mice or congenic controls. N= 6 independent replicates.

(L) Multistep viral growth curve analysis (MOI 0.1) in primary cortical neurons isolated from Ripk3-2xFV^fl/fl Mox2-Cre⁺ mice and Cre⁻ littermate controls. Prior to infection, cultures were pretreated (2h) with AP1 or DMSO vehicle. N= 4 independent replicates.

*p<0.05, **p<0.01, ***p<0.001. Error bars represent SEM. Dotted lines indicate limits of detection. All data are pooled from two or three independent experiments. See also Figure S2.