Fig. 5: Drosophila Tao activity is regulated by poly(I:C) binding and human TAOK2 affects ISG expression.
a THP-1 cell lysate was incubated with poly(I:C) or poly(C) agarose beads and input proteins or co-precipitating proteins were analyzed by western blotting against the indicated proteins. b Fluorescence quenching assay showing fluorescence elicited from FITC-tagged poly(I:C) in presence of increasing concentrations of dTao. Shown are the mean fluorescence intensity ±SD of three measurements. The indicated Kd was determined using Affinity Analysis v2.2.4 (NanoTemper Technologies). c Increasing amounts of dTao (50 and 200 ng/ml) were incubated or not with 0.3 mg/ml poly(I:C) and kinase activity was measured by a luminescence-based ATP consumption assay. Bars show the mean of three independent measurements ±SD. ****p < 0.0001, ***p < 0.001, ns p > 0.05 (Two-way ANOVA with Šídák’s multiple comparison test). AU arbitrary units. Data presented in (a, b, c) is representative of at least three independent experiments. (d, e, f) THP-1 KO or nontargeted control cells were seeded and infected with SFV (MOI 10) for 24 h and analyzed for proteome expression. Volcano plots show protein expression patterns of SFV-infected TAOK1 (e), TAOK2 (d), and TAOK3 (f) KO cells as compared to controls. Proteins with significantly different expression patterns are highlighted in blue, proteins belonging to the GO Term “cellular response to type-I interferon” are marked in orange and viral proteins in green. Data presented in (d, e, f) is averaged across four biological repeats; a two-sided Student’s t-test (S0 = 0.1, permutation-based FDR <0.05) was used to assess the significance. g, h THP-1 KO and nontargeted control cells (green: control, orange: TAOK1 KO, blue: TAOK2 KO, purple: TAOK3 KO, red: STAT1 KO) were seeded and infected with SFV (MOI 1) or left uninfected (Mock) for 24 h and analyzed for SFV RNA and MX1 mRNA transcript levels by RT-qPCR. Shown are the transcript levels normalized to the expression of the housekeeping gene GAPDH for four independent repeats expressed as fold change compared to the averaged biological repeats of the control. Error bars show the mean ± SD. ****p < 0.0001 (One-way ANOVA (g) or two-way ANOVA (h) both with Šídák’s multiple comparison test). ns not significant. Data presented in (g, h) is representative of three independent experiments. Source data are provided as a Source Data file.
