Inhibition of mTORC1 Enhances the Translation of Chikungunya Proteins via the Activation of the MnK/eIF4E Pathway
Fig 5
Enhanced protein translation during mTORC1 inhibition is specific for viral proteins.
(A) MEFs were infected with CHIKV (MOI = 1) in the presence of TORISEL for 24h (0.1 mg/ml) and labeled with puromycin (Puro) as detailed in the Methods section. Western blot was performed using anti-puromycin (clone 12D10), anti-E1/2 and anti-GAPD antibodies. Similar results were observed in three independent experiments. (B) HFF, HeLA or primary MEFs were infected with CHIKV (MOI = 0.1) in the presence of Rapamycin (100 nM) or TORISEL (0.1 mg/ml). Extracellular viral titers were determined 24h p.i. and results expressed as TCID50/ml. Bars indicate mean values ±SEM from four independent experiments. (C) MEFs were infected with CHIKV (MOI = 1), Sindbis expressing GFP (SINV; MOI = 0.1) or influenza A (Flu, 250 HAU106) for 24 h. The staining of E2, GFP and M2 proteins were analyzed using flow cytometry to quantify infection. Bars indicate mean values ±SEM from three independent experiments. Ø, control; T, TORISEL. Student’s test **, P < 0.05.