TRAF6 is not required for K13-induced NF-κB activation.

<p><b>A... </b><i>TRAF6^+/+</i> and <i>TRAF6^−/−</i> MEFs were transfected with a control vector or vector encoding K13 along with an NF-κB-Luc construct (75 ng/well) and a Renilla reporter construct (75 ng/well, normalization control) using Lipofectamine 2000-mediated transaction. The luciferase reporter assay was performed 48 hours post transfection essentially as described in “<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036601#s2" target="_blank">Materials and Methods</a>” section. Asterisks (*) indicate significance at levels of p≤0.05. <b>B... </b><i>TRAF6^+/+</i> and <i>TRAF6^−/−</i> MEFs were transfected with a control vector or a vector encoding K13-ER^TAM along with an NF-κB-Luc construct and a Renilla reporter construct. The transfected cells were subsequently treated with 4OHT (20 nM) for 48 hours and the reporter assays performed as described above. The values shown are averages (mean±SE) of one representative experiment out of three in which each transfection was performed in duplicate. Asterisks (*) indicate significance at levels of p≤0.05. <b>C... </b><i>TRAF6^+/+</i> and <i>TRAF6^−/−</i> MEF cells expressing an empty vector or the FLAG-tagged K13-ER^TAM treated with 4OHT were examined by immunoblot analysis for upregulation of A20. Tubulin was used as a loading control.