Fig. 6From: TNF-α enhances Th9 cell differentiation and antitumor immunity via TNFR2-dependent pathwaysTNF-α enhances Th9 cell differentiation through NF-κB signaling pathway. (a) Naïve CD4+ T cells were cultured under Th9 polarizing conditions with or without addition of TNF-α for 3 h. Western-blots examined the protein levels of phosphorylated IKKα/β (p-IKKα/β) and IκB-α in cells. β-actin was used as a loading control. (b) 293 T cells were transiently transfected with vectors contained Il9 promoter or empty vector, followed by transfecting with vectors expressing the indicated NF-κB molecules. Luciferase reporter assay showed NF-κB-dependent activation of Il9 promoter in 293 T cells. (c, d) Naïve CD4+ T cells were cultured under Th9 polarizing conditions in the presence or absence of TNF-α with or without (DMSO) the addition of NF-κB inhibitor bortizomib (Bor) for 3 days. IL-9 expression was examined by qPCR (c) and ELISA (d). Data are representative of three (a) independent experiments or presented as mean ± SD of three (b-d) independent experiments. *P < 0.05Back to article page