Skip to main content
. Author manuscript; available in PMC: 2014 Jan 20.
Published in final edited form as: Virology. 2012 Oct 12;435(2):415–424. doi: 10.1016/j.virol.2012.09.031

Fig. 6. Host general transcriptional suppression by RVFV MP-12 mutants.

Fig. 6

(A) Incorporation of uridine analog 5-ethynyluridine (EU) into nascent RNA is tested in MEF/wt cells. MEF/wt cells were mock-infected or infected with MP-12, rMP12-C13type, rMP12-NSsR173A, or rMP12-mPKRN167 at an moi of 3, and 1 mM EU was added into the culture at 16 hpi for 1 hour. Then, cells were fixed and stained with Alexa Fluor 594-coupled azide (red). The cells were further stained with anti-RVFV antibodies and Alexa Fluor 488-conjugated anti-mouse IgG (green). As a control, mock-infected cells were co-treated with actinomycin D (ActD) at 5 µg/ml with 1 mM EU for 1 hour at 16 hpi. (B) Fluorescence-activated cell sorting (FACS) analysis was performed in 293 cells. 293 cells were mock-infected or infected with MP-12, rMP12-C13type, or rMP12-NSsR173A at an moi of 3 and treated with 0.5 mM EU at 8 hpi for 1 hour. The control cells were co-treated with ActD (5 µg/ml) at 8 hpi for 1 hour. Incorporated EU was stained with Alexa Fluor 647-azide, and viral antigens were stained with anti-RVFV antibodies. Cells were then analyzed by flow cytometry on the LSRII Fortessa.