Extended Data Fig. 3: Ubiquitination of E in tissues from infected mice.

Tissues from testis (a), and brain (b) from mock or ZIKV-WT infected A129 mice were collected at day 8 post-infection. Tissues were homogenized and 200 μg of total input protein was used for immunoprecipitation (IP) of E using 4G2 antibody or an IgG control. Ubiquitination of WT-E was detected with anti-Ub antibody by immunoblot (IB). IPs shown are from mixed tissue lysates from 3 different mice. c-d, A129 mice (male and females) were mock treated (5 mice) or infected with ZIKV E-WT, ZIKV E-K38R or ZIKV E-K281R (1×10⁴ PFU, 9 mice per group, combined from 2 independent experiments). Weight loss and survival is shown in Fig 1e–f. c, serum titers (viremia), were determined at day 2 p.i. by plaque assay, after blood collection from 6 mice for ZIKV E-WT and K38R, and 7 mice for ZIKV E-K38R. d, virus titers (at day 8 p.i.) in brain (14 mice for ZIKV E- WT, and 9 mice for ZIKV E- K38R and ZIKV E- K281R), testis (6 mice/each gorup), and eye (14 mice for ZIKV E-WT, and 9 mice for ZIKV E-K38R and E-K281R). Unpaired, t-test, two-sided, *p < 0.05, **p < 0.01.