Fig. 6: AKIP1-PKA-CREB1 potentiates trVLPs and EBOV replication.

a, b WT and AKIP1^−/− (KO) HepG2 cells were transfected with EBOV minigenome (p0 and p1) for 96 h. Cells were lysed and the amounts of trVLPs in the cells were determined by luciferase activity assay (a). vRNA was quantified at the indicated times using qRT-PCR (b). Differences between the two groups were evaluated using the two-sided unpaired Student’s t-test. Data were presented as mean ± s.e.m. (**P < 0.01; ***P < 0.001). c, d HepG2 cells were transfected with the EBOV minigenome (p0 and p1) and treated with FSK (25 μM), H89 (10 μM), or 666-15 (1 μM) for 48 h. The amounts of trVLPs were determined by luciferase activity assay. Differences between the two groups were evaluated using the two-sided unpaired Student’s t-test. Data are presented as mean ± s.e.m. (*P  <  0.05; ***P  <  0.001). e HepG2 cells were transfected with the EBOV minigenome (p1) for 24 h or infected with live EBOV (MOI = 0.1). Then, the cells were treated with the indicated concentration of 666-15 or T-705 as a control for 24 h (for trVLP) and 96 h (for live EBOV infection). The amount of trVLPs was determined by luciferase activity assay, the live EBOV in the supernatant was titered by TCID₅₀ assay and the cytotoxicity of 666-15 toward HepG2 cells was determined by performing CCK-8 assay. f WT and AKIP1^−/− HepG2 cells infected with live EBOV (MOI = 1) were treated with 10 μM H89 or 1 μM 666-15 for 96 h. vRNA levels in the cells were quantified by qRT-PCR. Differences between the two groups were evaluated using the two-sided unpaired Student’s t-test. Data were presented as mean ± s.e.m. ***P < 0.001. g HepG2 cells infected with live EBOV (MOI = 0.1) were treated with 10 μM H89, 1 μM 666-15, or 50 μM T-705. The viruses in the supernatants of cell culture were quantified by reverse transcription and qRT-PCR. h HepG2 cells infected with live EBOV (MOI = 1) were treated with 1 μM 666-15. The cell culture supernatants were collected at the indicated time points, and the viral titers were quantified as TCID₅₀ with plaque assay. Differences between the two groups were evaluated using the two-sided unpaired Student’s t-test. The data from three independent experiments are presented as the means ± s.e.m. (*P < 0.05).