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[Preprint]. 2023 Jul 15:2023.07.14.549057. [Version 1] doi: 10.1101/2023.07.14.549057

Figure 1. EBOV VP35 protein interacts with Ub non-covalently.

Figure 1.

A) Whole Cell extracts (WCE) from HEK293T cells transfected with Flag-VP35 (VP35) and HA-Ub wild type (WT) or HA-Ub DGG (cannot conjugate proteins), were used for HA immunoprecipitation (IP) under non-denaturing conditions (RIPA washes), followed by immunoblot (IB). B) Purified recombinant K48 or K63 polyUb chains (mix of 2–7 Ub chains), were mixed in vitro with Flag-VP35, followed by Flag IP. Interacting proteins were eluted with Flag peptide. C-D) Experiments performed as in (B) but using the C-terminal IID domain of VP35 (C), or the VP35 K309R or K309G mutants, which are not covalently ubiquitinated (D). * No ubiquitinated VP35, possibly phosphorylation.