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. 2019 Jun 21;15(6):e1007875. doi: 10.1371/journal.ppat.1007875

Fig 5. Effect of suppressing specific host gene function on viral gene expression.

Fig 5

(A) GFP expression of a rVSV-eGFP reporter virus in uninfected, vehicle-treated, or 2.5 μM 17-DMAG treated HeLa cells. A representative histogram of GFP intensity is shown to the right, and the mean fluorescence intensity (MFI) of live cells is shown below. MFI is normalized to vehicle, and error bars denote the standard deviation from the mean of 3 independent replicates. (B) VSV gene expression in eIF3a depleted HeLa cells. A representative histogram of fluorescence intensity is shown to the right and the MFI below. Error bars denote standard deviation from the mean from three independent replicates. Luciferase expression driven by rVSV-LUC (Firefly Luciferase) or a cellular reporter driving expression of Renilla Luciferase (pRL-CMV) in eIF3a depleted cells. Luciferase expression is presented as the percent of a non-targeting siRNA control, and error bars denote the standard deviation from 3 independent replicates. Metabolic labeling of eIF3a depleted HeLa cells infected with wild-type VSV. The position of viral proteins is noted to the right. Presented is a representative gel from two independent replicates. (C) VSV gene expression and replication in DDIT4 depleted cells. A representative histogram of eGFP expression is shown along with the MFI of cells normalized to a non-targeting siRNA control. Error bars denote the standard deviation from the mean of three independent replicates. For metabolic labeling a representative gel of two independent replicates is presented. Kinetics of viral replication were measured by titration of yields at various times post infection of siRNA treated HeLa cells. Error bars denote the standard deviation from the mean of 3 independent replicates. (D) Immunoprecipitation of cellular proteins synthesized post-infection with VSVWT. Shown is a representative gel from two independent replicates. A quantitative analysis of the HSP70 and YBX1 bands is shown in the bottom two panels, error bars denote the standard deviation from two independent replicates.