Effect of UBB⁺¹ on the NF-κB and JNK pathway.

<p>(A) CRT-MG cells stably expressing pEGFP or pEGFP-UBB⁺¹ were treated with TNF-α (10 µg/L) for the indicated time periods. Cell lysates were examined by immunoblot analysis with antibodies to phospho-IKK, IKK, phospho-IκBα, IκBα and β-actin. (B) The relative amount of phosphorylated IKK, phosphorylated IκBα and total IκBα was plotted over the time upon TNF-α treatment in pEGFP-UBB⁺¹ stable cells (dotted line) and pEGFP stable cells (solid line). Ratiometric measurement of each blot was normalized by the value of β-actin. Representative of four independent experiments. (C) Cells stably expressing pEGFP or pEGFP-UBB⁺¹ were treated with TNF-α (10 µg/L) for the indicated time periods. Cell lysates were examined by immunoblot analysis with antibodies to phospho-MKK4, MKK4, phospho-JNK, JNK, phospho-c-Jun, c-Jun, phospho-ERK1/2, ERK1/2 and β-actin. (D) The relative amount of phosphorylated MKK4, phosphorylated JNK and phosphorylated c-Jun was plotted over the time upon TNF-α treatment in pEGFP-UBB⁺¹ stable cells (dotted line) and pEGFP stable cells (solid line). Ratiometric measurement of each blot was normalized by the value of β-actin. Representative of four independent experiments.