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. 2010 Dec 7;286(6):4912–4921. doi: 10.1074/jbc.M110.199729

FIGURE 5.

FIGURE 5.

Spliced XBP1 blocked TNF-α induced IKK activation through inhibition of IRE phosphorylation and up-regulation of GRP78. A, HREC were infected with Ad-XBP1S or Ad-GFP for 48 h, followed by treatment with 10 ng/ml TNF-α for up to 60 min. Phosphorylation of IKKα/β at Ser176/180, IκB-α at Ser32, and NF-κB at Ser536 were detected by Western blot analysis. Total IKKα, IKKβ, IκB-α, and NF-κB were also determined. p, phosphorylated; t, total. Representative results were from three independent experiments. B–D, HREC were infected with Ad-XBP1S or Ad-GFP as described and treated with 10 ng/ml TNF-α for indicated time periods. Expression of IκB-β (B), total and phospho-IRE1α (C), and GRP78 (D) were determined by Western blot analysis. E, HREC were pretreated with tunicamycin (TM) followed by exposure to TNF-α for 4 and 24 h. Expression of GRP78 were determined by Western blot analysis. **, p < 0.01 versus Ad-GFP (D) or control (E).