Fig. 7: EBOV VP35 promotes the transcription of CREB1-directed coagulation-related genes.

a HepG2 cells cotransfected with pGL-CRE-Luc, pRL-TK, and the indicated amounts of Flag-VP35 were treated with or without 25 μM FSK for 4 h. The luciferase activity of the cell lysates was analyzed. Differences between the two groups were evaluated using a two-sided unpaired Student’s t-test. The mean ± s.e.m. from three independent assays is presented (n = 3; ***P < 0.001). b, c HUVECs (b) and WT or AKIP1^−/− HepG2 cells (c) infected with Ad-VP35 (MOI = 10) (b) or live EBOV (MOI = 1) (c) for 48 h were treated with or without 10 μM H89 or 1 μM 666-15 for another 24 h. THBD and SERPINB2 mRNA levels were determined 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. (n = 3; ***P  <  0.001). d WT or Akip1^−/− mice were intravenously injected with Ad-VP35 or Ad-null (2 × 10⁹ PFU) twice at an interval of 24 h. Six days post the first infection, the liver tissues were analyzed by immunohistochemistry staining with an anti-Thrombomodulin (TM) antibody. e, f Mice were infected with Ad-VP35 or Ad-null (3 × 10⁹ PFU), treated with 666-15 (2 mg/kg) or solvent and then challenged with or without LPS. The tail bleeding time was determined (n = 8) (e), and a mouse survival curve is shown in (f) (n = 9). Differences between the two groups were evaluated using the two-sided unpaired Student’s t-test (e). Survival curves were analyzed by log-rank test (f). All data from two independent experiments are presented as the means ± s.e.m. (ns not significant; *P < 0.05; **P < 0.01).