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. 2018 Jun 11;9:1327. doi: 10.3389/fimmu.2018.01327

Figure 8.

Figure 8

Involvement of Danio rerio DDX41 (DrDDX41) in Danio rerio STING (DrSTING)–Danio rerio STAT6 (DrSTAT6)-mediated Danio rerio CCL20 (DrCCL20) expression. (A) Quantitative RT-PCR (Q-RT-PCR) analysis of HsCCL20 in HEK293T cells transfected with increasing amounts (0, 200, and 400 ng/mL) of expression vectors for DrDDX41 or DrSTING individually or expression vector for DrSTING (200 ng/mL) together with increasing concentrations (0, 200, and 400 ng/mL) of expression vectors for DrDDX41 and stimulated with poly(dA:dT) (1 µg/mL) for 6 h. Data are representative of three independent experiments as mean ± SD (*p < 0.05; **p < 0.01). (B) Q-RT-PCR analysis of HsCCL20 in HEK293 cells transfected with increasing amounts (0, 200, and 400 ng/mL) of expression vectors for DrDDX41, DrSTING or DrSTAT6 individually or expression vector for DrSTAT6 (200 ng/mL) together with increasing concentrations (0, 200, and 400 ng/mL) of expression vectors for DrDDX41 or DrSTING and stimulated with poly(dA:dT) (1 µg/mL) for 6 h. Data are representative of three independent experiments as mean ± SD (*p < 0.05; **p < 0.01). The bands in the upper-left denote the western blot analysis of the expression of DrDDX41, DrSTING, and DrSTAT6 in HEK293 cells transfected with increasing amounts (0, 200, and 400 ng/mL) of corresponding vectors. (C) DrSTING coimmunoprecipitates with DrSTAT6 through the mediation of DrDDX41 under poly(dA:dT) stimulation. (D) Q-RT-PCR analysis of DrCCL20 mRNA levels in 24 hours post microinjection embryos microinjected with standard MO and PBS (US) or standard MO (Ctrl), DrDDX41/DrSTING/DrSTAT6-MO, the targeted-MOs together with their corresponding mRNAs, and poly(dA:dT) (100 pg/embryo). Data are representative of three independent experiments as mean ± SD (*p < 0.05; **p < 0.01). Standard loading was indicated by β-actin expression. (E) Q-RT-PCR analysis of DrCCL20 mRNA levels in embryos microinjected with different combinations of DrDDX41-MO (DrD-MO), DrSTING-MO (DrS-MO), DrSTAT6-MO (DrS6-MO), and their mRNAs (mR) for rescue, and poly(dA:dT) (100 pg/embryo). Each of the MOs and mRNAs was administered at 4 ng/embryo and 200 pg/embryo in each group. Data are representative of three independent experiments as mean ± SD (*p < 0.05; **p < 0.01). Standard loading was indicated by β-actin expression.