Figure 6
Effects of recombinant Ae. albopictus ADA on DENV-2 replication. Raw264.7 cells (A) and human THP-1 cells (B) were treated with DENV-2 (MOI = 0.25) alone or in combination with various doses (0.02, 0.2, and 2 μg/mL) of recombinant Ae. albopictus ADA. At various time points (6 h,12 h, and 24 h) following the stimulation, RNA was isolated from the cells, cDNA was generated, and qRT-PCR was performed to detect the genomes of the DENV envelope. Raw 264.7 cells treated with DENV-2 alone or in combination with recombinant Ae. albopictus ADA (2 μg/mL) and untreated group (Medium) were stained for dsRNA detection (J2 antibody); a flow cytometry (C) was applied to analyze the J2 fluorescence signal: representative result obtained from the flow cytometry analysis (left panel) and quantification of mean fluorescence intensity (MFI) (right panel) with n = 3. The data obtained from the analysis of Raw 264.7 cells were normalized to the mouse GAPDH using the ΔΔCT method and are presented as percentages of the average ΔΔCT value of the cells treated with DENV-2 alone; human THP-1 cells were normalized to human actin using the ΔΔCT method and are presented as percentages of the average ΔΔCT value of the cells treated with DENV-2 alone. A nonparametric Mann–Whitney test was applied for the statistical analysis. Labels: ns, insignificant (p > 0.05); *p < 0.05; **p < 0.01; ***p < 0.001. The data are representative of three independent experiments.
