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. Author manuscript; available in PMC: 2021 Jan 8.
Published in final edited form as: Nature. 2020 Jul 8;585(7825):414–419. doi: 10.1038/s41586-020-2457-8

Fig. 5. Ubiquitination of ZIKV E promotes binding to the Tim-1 receptor.

Fig. 5.

a, WCE from HEK293Ts transfected with empty vector (EV), HA-E WT and Flag-TIM-1 were used for IP with anti-Flag beads. b, Interaction of ZIKV E-WT but not ZIKV-E K38R viral particles with TIM-1. Ectopically expressed TIM-1 was isolated using anti-Flag beads. After washes, viruses were incubated with the TIM-1. c, Interaction of K63-, but not K48-linked polyUb, with TIM-1 in the absence of E, coIP assay. d, WT was bound to TIM-1 containing beads (as in b). Increasing amounts of purified K63-Ub or K48-Ub chains were added. Representatives of 2 independent experiments (a-d). e, Virus attachment (viral RNA, qPCR) after Tim-1 knockdown in JEG-3 (n=2, mean +/−SE, One-Way ANOVA, multiple comparison, Tukey correction, ****p<0.0001, *p<0.05, NS p>0.05). f, Titers in infected Tim-1−/− mice, n=5/group, mean +/−SE, Two-Way ANOVA multiple comparison, Tukey correction. ****p<0.0001, NS p>0.05.