Fig. 3.

PBA-induced HDPs gene expression activates the NF-κB signaling pathway. (a) qRT-PCR analysis of p50 and p65 transcription in cells treated with 8 mM PBA for 24 h. (b) Immunoblot analysis of the phospho-NF-κB p65 protein and the IκB-α protein in cells pretreated or not for 0–24 h with 8 mM PBA. (c) IPEC J2 cells were co-transfected with the pNF-kB-Luc and the internal-control phRL-TK plasmids for 6 h, and were treated with PBA or LPS for 24 h. The cells were then harvested and analyzed by luciferase reporter assays. (d) The IPEC J2 cells were transfected with TLR2/4 siRNA to specifically silence TLR2/4 for 6 h and were then challenged with PBA at 8 mM for 24 h, and a Western blot was used to assess the protein levels of IκB-α and phospho-p65. A densitometric analysis of the IκB-α and phospho-p65 protein levels is represented as the mean ± SD from six independent experiments. Letters with different superscripts are significantly different at P < 0.01, compared with the vehicle.