Journal: iScience
Article Title: FDA-approved Abl/EGFR/PDGFR kinase inhibitors show potent efficacy against pandemic and seasonal influenza A virus infections of human lung explants
doi: 10.1016/j.isci.2023.106309
Figure Lengend Snippet: NRTKIs affect IAV RNA replication (A) A549 cells were transfected with pPOLI-358-FFluc and pmaxGFP plasmids. At 24 hpt, cells were infected with NL09 or NL11 at MOI = 1 +/− indicated NRTKIs at [0.5x or 1x] max concentrations. At 48 hpt (24 hpi), luciferase activity was measured and normalized to GFP expression (MFI). (B) GFP-normalized polymerase activity of untreated NL09-or NL11-infected cells is shown. (C) A549 cells were transfected with pPOLI-358-FFluc and pmaxGFP plasmids and co-transfected with NL09 or NL03-minigenome plasmids. At 6 hpt, NRTKIs were added to the medium. At 30 hpt (24 h of treatment), luciferase activity was measured and normalized to GFP MFI. Bars indicate values relative to untreated cells normalized to GFP. (D) GFP-normalized polymerase activity of untreated NL09 or NL03 minigenome transfected cells is shown. Triplicate measurements from triplicate samples (n = 3); error bars indicate ±standard deviation (SD). ∗, p < 0.05; ∗∗, p < 0.01; ∗∗∗, p < 0.001; ∗∗∗∗, p < 0.0001. p-values determined by Brown-Forsythe and Welsh ANOVA compared to untreated.
Article Snippet: Semi-confluent (∼70–80%) A549 cells (8×10 4 cells in 24-well plates) were transfected using Lipofectamine LTX with the pPOLI-358-FFLuc reporter plasmid, which encodes a firefly luciferase gene under control of the viral nucleoprotein (NP) promoter (kindly provided by Megan Shaw) , , ; the Lonza pmaxGFP™ expression vector, was used as a transfection control.
Techniques: Transfection, Infection, Luciferase, Activity Assay, Expressing, Standard Deviation
Lonza manufactures this product 
![NRTKIs affect IAV RNA replication (A) A549 cells were transfected with pPOLI-358-FFluc and <t>pmaxGFP</t> plasmids. At 24 hpt, cells were infected with NL09 or NL11 at MOI = 1 +/− indicated NRTKIs at [0.5x or 1x] max concentrations. At 48 hpt (24 hpi), luciferase activity was measured and normalized to GFP expression (MFI). (B) GFP-normalized polymerase activity of untreated NL09-or NL11-infected cells is shown. (C) A549 cells were transfected with pPOLI-358-FFluc and pmaxGFP plasmids and co-transfected with NL09 or NL03-minigenome plasmids. At 6 hpt, NRTKIs were added to the medium. At 30 hpt (24 h of treatment), luciferase activity was measured and normalized to GFP MFI. Bars indicate values relative to untreated cells normalized to GFP. (D) GFP-normalized polymerase activity of untreated NL09 or NL03 minigenome transfected cells is shown. Triplicate measurements from triplicate samples (n = 3); error bars indicate ±standard deviation (SD). ∗, p < 0.05; ∗∗, p < 0.01; ∗∗∗, p < 0.001; ∗∗∗∗, p < 0.0001. p-values determined by Brown-Forsythe and Welsh ANOVA compared to untreated.](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_4449/pmc10034449/pmc10034449__gr6.jpg)