FIGURE 7.

Knockdown of NF-κB (p65) reduces IL-9 expression. In vitro differentiated primary Th9 cells were transfected with siRNA against NF-κB (p65) or non-relevant siRNA (mock). A, the knockdown efficiency of p65 was confirmed by immunoblotting the respective cell lysates with antibodies against p65 and lamin B (control). B, relative level of IL-9 transcript in the mock- and p65 siRNA-transfected cells were analyzed by qRT-PCR and represented relative to the expression level of the housekeeping gene hypoxanthine-guanine phosphoribosyltransferase. C, p65 siRNA- or mock-transfected Th9 cells were stimulated for 24 h with PMA/ionomycin, and the IL-9 protein levels in the culture supernatant were measured by ELISA. The data shown are expressed as mean ± S.E. (error bars) (n = 3); *, p < 0.05; **, p < 0.01.