Figure 5
Recombinant Ae. albopictus ADA activates the NF-κB pathway by targeting Takinib. (A) Immunoblot analysis of phosphorylated IκBα(p-IκBα), total proteins of IκBα(IκBα), phosphorylated p65(p-p65), and p65 in human THP-1 cells from untreated (Medium) or treated with ADA (2 μg /mL) groups at various time points (15 min, 30 min, and 1 h). (B) Confocal microscopy of p65 in mouse Raw264.7 cells from untreated (Medium) or treated with ADA (2 μg /mL) groups at 1 h. (C) Human THP-1 cells treated with the inhibitors, Takinib (10 μM) or DMSO, for 3 h before being stimulated with ADA (2 μg/mL) at various time points (15 min, 30 min, and 1 h); p-IκBα, IκBα, p-p65and p65 were analyzed via Immunoblot assay. (D) Human THP-1 cells were treated with the inhibitors, Takinib (10 μM) or DMSO, for 3 h before stimulation with ADA (2 μg/mL) for 24 h; RNA was then isolated from the cells, cDNA was generated, and qRT-PCR was performed to measure the mRNA level of IL-1β, IL-6, and TNF-α. A nonparametric Mann–Whitney test was applied for the statistical analysis. *p < 0.05; **p < 0.01. The data are representative of two independent experiments.
