Smad phosphorylation is dependent on RVFV replication and NSs expression.

<p>A) Analysis of HSAECs infected with MP12, ZH548 or their UV-inactivated (UVin) controls at 9 and 18hpi. Protein lysates were analyzed by western blot for pSmad1/5/9 MH2, pSmad1/5 MH2, pSmad2 MH2, pSmad2 L, RVFV NP, actin as well as total protein levels for Smad1 and Smad2. M: Mock; MP: MP12; ZH: ZH548. B) Smad phosphorylation in HSAECs infected with increasing MOIs (0.1, 0.5, 1, and 5) of MP12 virus at 9hpi was examined as described in panel A. C) Analysis of Vero cells infected at an MOI 3 with MP12 or MP12 ΔNSs. With the exception of pSmad1/5/9 MH2, protein lysates were analyzed by western blot as described in panel A. D) Vero cells transfected with media alone, pcDNA3.1 vector (2.5 μg), or pcDNA3.1-Flag-NSs (2.5 μg) were harvested at 24 hours post transfection. Protein lysates were analyzed by western blot for pSmad1/5 MH2, pSmad2 MH2, pSmad2 L, Flag-NSs as well as total protein levels for Smad1 and Smad2.</p>