Fig. 6: TAOK2 is required for IFN induction in infected and poly(I:C) stimulated cells. | Nature Communications

Fig. 6: TAOK2 is required for IFN induction in infected and poly(I:C) stimulated cells.

From: Cross-species analysis of viral nucleic acid interacting proteins identifies TAOKs as innate immune regulators

Fig. 6

a THP-1 cells were infected with SFV-mCherry (MOI 5) and red fluorescence intensity was measured every 3 h using an Incucyte S3 live-cell imaging system. The line diagrams show the mean integrated red intensity/cell confluence per image (RCU) ± SD (y-axis) over time (x-axis). b Nontargeted control (Control), TAOK2 KO, or STAT1 KO THP-1 cells were left unstimulated (Mock) or stimulated with SFV (MOI 5) or IFN-α B/D (1000 units/mL) for 24 h and used for western blotting against the indicated proteins. c Nontargeted control (Control), TAOK2 KO, or STAT1 KO A549-IFIT1-eGFP cells were infected with SFV (MOI 5) and green fluorescence intensity was measured at the indicated time points using an Incucyte S3 live-cell imaging system. Mean green intensity/cell confluence per image (GCU) ± SD (y-axis) is shown over time (x-axis). d Representative fluorescence microscopy images of (c) 24 h after infection with SFV. e, f as (c) but transfected with poly(I:C) (2 µg/mL) (e) or stimulated with IFN-α B/D (1000 units/ml) (f). Control cells are colored in green, TAOK2 KO cells in blue, and STAT1 KO cells in red. g A549-IFIT1-eGFP cells were infected with SFV-mCherry (MOI 5) and simultaneously treated with the TAOK2 inhibitor RAF265 (500 nM, blue), BRAF inhibitor Dabrafenib (500 nM, red), or left untreated (green). Green (GCU) and red fluorescence (RCU) intensities were measured at the indicated time points using an Incucyte S3 live-cell imaging system. Mean green or red intensity/cell confluence per image (G/RCU) ± SD (y-axis) is shown over time (x-axis). h A549-ACE2 cells were treated with the TAOK2 inhibitor RAF265 (10 µM) and infected with SARS-COV-2-GFP (MOI 3) and accumulation of GFP was measured over time in an Incucyte S3 system. Line diagrams show the mean green intensity/cell confluence per image (GCU) ± SD (y-axis) over time (x-axis). Data presented in (a–h) is representative of three independent experiments and for each line diagram, the mean ± SD are plotted based on at least four biological repeats. ****p < 0.0001, ***p < 0.001, **p < 0.01, *p < 0.05, ns p > 0.05 (Repeated measurements two-way ANOVA with Šídák’s multiple comparison test to compare control versus treatment conditions). Source data are provided as a Source Data file.

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