Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Integrated phenotypic screening and chemical proteomics identifies ETF1 ligands that modulate viral translation and replication

doi: 10.1073/pnas.2524108123

Figure Lengend Snippet: Photo-stereoprobes inhibit additional viruses that use PRF. ( A ) Effect of active stereoprobe WX-02-31 and control stereoprobes on HIV-1 pseudovirus production. ( Top ) Schematic of the HIV-1 ribosomal frameshift sequence. ( Bottom ) HEK293T cells expressing the HIV-1 NL4-3 luciferase vector were treated with the indicated photo-stereoprobes (10 µM) and assessed for luciferase activity after 24 h. Data are average values ± SD for 2 to 5 independent experiments that each have three technical replicates. ( B ) Effect of active stereoprobe WX-02-31 and control stereoprobes on Sindbis virus (SINV) infection. ( Top ) Schematic of the SINV ribosomal frameshift sequence. ( Bottom ) HeLa cells were pretreated with indicated photo-stereoprobes (10 µM, 3 h), inoculated with SINV (MOI 1) in the presence of compounds, and viral supernatant titers were assessed after 48 hpi. Data are average values ± SD for three independent experiments that each have 2 to 3 technical replicates. ( C ) Effect of active stereoprobe WX-02-31 and control stereoprobes on porcine reproductive and respiratory syndrome virus (PRRSV) infection. ( Top ) Schematic of the PRRSV ribosomal frameshift sequence. ( Bottom ) MA-104 cells were pretreated with indicated photo-stereoprobes (10 µM, 3 h), inoculated with PRRSV-GFP (MOI 0.1) in the presence of compounds, and viral antigen levels were assessed by GFP expression after 24 hpi. Data are representative of two independent experiments that each have three technical replicates. ( D ) Effect of active stereoprobe WX-02-31 and control stereoprobes on lymphocytic choriomeningitis virus (LCMV) infection. ( Top ) Schematic of the LCMV genome. ( Bottom ) HeLa cells were pretreated with indicated photo-stereoprobes (10 µM, 3 h), inoculated with LCMV (clone 13, MOI 0.01) in the presence of compounds and viral supernatant titers were assessed after 48 hpi. Data are average values ± SD for two independent experiments that each have 2 to 3 technical replicates. ( E ) Effect of active stereoprobe WX-02-31 and control stereoprobes on influenza A virus (IAV) infection. ( Top ) Schematic of the IAV genome. ( Bottom ) Calu-3 cells were pretreated with indicated photo-stereoprobes (20 µM, 3 h), inoculated with IAV (MOI 0.005) in the presence of compounds, and viral antigen levels were assessed by flow cytometry after 48 hpi. Data are the average values ± SD for two independent experiments that each have three technical replicates. For ( A – C ), two-tailed unpaired t test comparing WX-02-31 versus DMSO or WX-02-51 groups: * P < 0.05, **** P < 0.0001.

Article Snippet: Porcine reproductive and respiratory system virus with GFP (PRRSV-GFP) was kindly provided by Jay Calvert (Zoetis) via Bob Rowland (University of Illinois) and propagated on MARC-145 cells ( ).

Techniques: Control, Sequencing, Expressing, Luciferase, Plasmid Preparation, Activity Assay, Virus, Infection, Flow Cytometry, Two Tailed Test