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. 2017 Jun 5;98(5):1058–1072. doi: 10.1099/jgv.0.000736

Fig. 7.

Fig. 7.

Virions and dense bodies (DBs) delivered miRNA to host cells may affect cellular functions, as shown by luciferase reporter assay. (a, b) HCMV-miR-US25-1-5p luciferase reporter assay – HEK-293 cells transfected with luciferase reporter constructs containing CCNE2 5′ untranslated region (CCNE2) or its mutated control (CCNE2-MUT) together with Renilla vector as internal control. (a) First luciferase experiment: on the transfected cells UV-irradiated virions, or DBs, or mock samples were added. (b) Second luciferase experiment: to further strengthen the data obtained from the first luciferase reporter assay experiments and improve the specificity of the assay. Virions and DBs were purified from the miR-US25 knockout (miR-US25KO) and its control wild-type (WT) virus stocks, and particles were UV irradiated and added to the HEK-293 cells transfected with CCNE2 or CCNE2-MUT luciferase reporters (the amount of particles was doubled in the second luciferase experiment compared to the first). In both the first and the second luciferase experiments as positive and negative miRNA controls hcmv-miR-US25-1-5p mimic or scramble miRNA mimics were co-transfected with the luciferase constructs, respectively. Cells were lysed 20 h later and luciferase activity was quantified. Firefly luciferase activity was normalized to Renilla luciferase; values are percentage mean±sem of three experiments performed in triplicate. P<0.05 was considered statistically significant. *P<0.05; **P<0.01; ***P<0.001.