Figure 6

Th9 immune response suppression following NF-κB gene knockdown. A: Efficiency of NF-κB interference using different siRNAs. The transcription of NF-κB was detected using qPCR by introducing different siRNA/NF-κB sequences (siRNA-1, siRNA-2, and siRNA-3) and nonspecific (ns) siRNA using RNAiMAX Transfection Reagent for 48 h. Only RNAiMAX Transfection Reagent was added to the blank group. The significance levels were set as follows: *p < 0.05, **p < 0.01, and ***p < 0.001, with “ns” denoting nonsignificance compared with the ns siRNA group. B: Efficiency of NF-κB interference with different concentrations of siRNA-1/NF-κB. Goat peripheral blood mononuclear cells (PBMCs) were treated with different concentrations of siRNA-1/NF-κB (0, 10, 20, 40, and 80 µg/mL) using RNAiMAX Transfection Reagent for 48 h. The significance levels were defined as ****p < 0.0001, with “ns” representing nonsignificance compared with the control (blank, 0 nM). Subsequently, goat PBMCs were incubated with 20 µg/mL rHcL6 for 24 h after the addition of 20 nM siRNA-1/NF-κB and ns siRNA using RNAiMAX Transfection Reagent. The control group was also treated with 20 µg/mL rHcL6 for 24 h after the addition of RNAiMAX Transfection Reagent. The significance levels were set at *p < 0.05, **p < 0.01, and ***p < 0.001, with “ns” indicating nonsignificance compared with the ns siRNA group. Data are representative of three independent experiments. C: Relative fold changes in GATA3, IRF4, PU.1, SMAD, STAT1, STAT6, STAT5 and NFAT transcription after NF-κB gene knockdown. D: Relative fold change in IL-9 transcription after NF-κB gene knockdown.