Fig. 4: EBOV VP35 promotes CREB1 phosphorylation at S133 in vitro and in vivo via AKIP1.
From: Ebola virus VP35 hijacks the PKA-CREB1 pathway for replication and pathogenesis by AKIP1 association
a HepG2 cells infected with Ad-VP35 or Ad-GFP (MOI = 10) for 48 h were treated with 25 μM FSK or 10 μM H89 for 4 h and then analyzed using immunoblotting. b HepG2 cells expressing GFP-VP35 or GFP were subjected to immunostaining with an anti-pCREB1(S133) antibody (red). c HepG2 cells infected with live EBOV (MOI = 10) for 72 h (upper panel) or transfected with EBOV minigenome (p0) for 48 h (lower panel) were subjected to immunostaining with the indicated antibodies. d HepG2 cells transfected with the AKIP1 siRNA or scrambled (Scr) siRNA were infected with Ad-VP35 or Ad-GFP (MOI = 10) for 36 h. Then, lysates were analyzed using immunoblotting. e Lysates of WT and AKIP1−/− (two independent clones, KO1 and KO2) HepG2 cells infected with Ad-VP35 or Ad-GFP (MOI = 10) were analyzed using immunoblotting. f C57BL/6 N mice were intravenously injected with Ad-VP35 or Ad-null (2 × 109 PFU) twice at an interval of 24 h. Three days after the first infection, the liver (upper panel) and lung (lower panel) tissues were analyzed using immunohistochemical staining with an anti-pCREB1 (S133) antibody. At least two independent replicates were performed in all experiments.
