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. Author manuscript; available in PMC: 2019 May 1.
Published in final edited form as: Virology. 2018 Mar 27;518:335–348. doi: 10.1016/j.virol.2018.03.016

Figure 1. Construction and confirmation of ECTVΔE3L.

Figure 1

(A) Uninfected w.t. BS-C-1 and BS-C-1+E3L cells were stained intracellularly for E3 protein and analyzed using flow cytometry. The positive gate was set based upon staining in the w.t. cells, which are negative for E3. (B) Representation (not drawn to scale) of the genome of w.t. ECTV in the region of the E3L gene (ECTV-043) and the approximate location where the GFP cassette was inserted to disrupt the coding region of E3L. The arrows depict the binding locations of PCR primers (P). (C and D) Using the primers shown in (B), PCR reactions were performed to confirm deletion of the predicted sequence from the E3L locus. The positive control (pos. ctrl.) reaction was carried out using primers (not depicted) to amplify a shared sequence located elsewhere in the genome. (E) BS-C-1 cells were infected (MOI=5) for 18 hrs prior to isolating total protein. A Western blot was performed to detect E3 from these cellular extracts. Separate lanes of the SDS-PAGE gel were loaded with equal amounts of total protein to detect actin, which was used as a loading control.