Figure 5

Th9 immune response suppression following STAT6 gene knockdown. A: Efficiency of STAT6 interference using different siRNAs. The transcription of STAT6 was detected using qPCR by introducing different siRNA/STAT6 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 and ****p < 0.0001, with “ns” denoting nonsignificance compared with the ns siRNA group. B: Efficiency of STAT6 interference with different concentrations of siRNA-2/STAT6. Goat peripheral blood mononuclear cells (PBMCs) were treated with different concentrations of siRNA-2/STAT6 (0, 10, 20, 40, and 80 µg/mL) using RNAiMAX Transfection Reagent for 48 h. The significance levels were defined as *p < 0.05, **p < 0.01, and ***p < 0.001, 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-2/STAT6 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 < 0.01, 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, STAT5, NFAT, and NF-ĸB transcription after STAT6 gene knockdown. D: Relative fold change in IL-9 transcription after STAT6 gene knockdown.