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. 2014 Dec 11;9(12):e114791. doi: 10.1371/journal.pone.0114791

Figure 5. Effect of LTβR siRNA on IL-8 production by BEAS-2B cells.

Figure 5

BEAS-2B cells were transfected with LTβR siRNA to knock down the receptor. (A) We purchased two types of siRNA (#1 and #2) and transfected them into BEAS-2B cells. We evaluated the knockdown efficacy of each siRNA 72 h later by qRT-PCR and western blotting. Lipofectamine reagent and the negative control siRNA (NC siRNA) did not affect LTβR mRNA expression, but siRNA#1 and #2 both significantly inhibited LTβR mRNA. (B) BEAS-2B cells were transfected with siRNA, and 72 h later they were stimulated with 50 ng/ml LIGHT. The IL-8 and IL-6 concentrations in the cell supernatants were determined by ELISA 24 h after stimulation. Both siRNA#1 and siRNA#2 significantly inhibited IL-8 and IL-6 production by BEAS-2B cells. n = 4 separate experiments. **: p<0.01. (C) BEAS-2B cells were pre-incubated with LTβR blocking antibody before stimulation with 50 ng/ml LIGHT. The blocking antibody for LTβR significantly attenuated both IL-8 mRNA expression and IL-8 production. n = 3 separate experiments. **: p<0.01.