Extended Data Fig. 3: Ubiquitination of E in tissues from infected mice.
From: Envelope protein ubiquitination drives entry and pathogenesis of Zika virus
a, b, Tissues from testis (a) and brain (b) from mock-infected or wild-type-ZIKV-infected A129 mice were collected at day 8 after infection. Tissues were homogenized and 200 μg of total input protein was used for immunoprecipitation of E using 4G2 antibody or an IgG control. Ubiquitination of wild-type E was detected with anti-ubiquitin antibody by immunoblot. Immunoprecipitations shown are from mixed tissue lysates from three different mice. c, d, A129 mice (male and females) were mock-treated (5 mice) or infected with wild-type ZIKV, E(K38R)- or E(K281R)-mutant ZIKV (1 × 104 PFU, 9 mice per group, combined from 2 independent experiments). Weight loss and survival is shown in Fig. 1e, f. c, Serum titres (viraemia), were determined at day 2 after infection by plaque assay, after blood collection from 6 mice for wild-type and E(K281R)-mutant ZIKV, and 7 mice for E(K38R)-mutant ZIKV. d, Virus titres (at day 8 after infection) in brain (14 mice for wild-type ZIKV, and 9 mice for E(K38R)- and E(K281R)-mutant ZIKV), testis (6 mice per group) and eye (14 mice for wild-type ZIKV, and 9 mice for E(K38R)- and E(K281R)- ZIKV). Unpaired, t-test, two-sided, *P < 0.05, **P < 0.01.
