Nf κb pathway sampler kit

The following antibodies were used: NF-κB Pathway Sampler Kit (#9936) (New England BioLabs, Frankfurt/Main, Germany), anti-pSer-43-NEMO (Abgent, Heidelberg, Germany), phospho-specific anti-pSer-43-NEMO (Eurogentec, Köln, Germany), anti-Ku70 (#sc-1487), anti-DNA-PKcs (#sc-5282) (Santa Cruz Biotechnology, Heidelberg, Germany), anti-Ku70/Ku80 (Abcam, Cambridge, UK), anti-ubiquitin from DAKO (Hamburg, Germany), anti-IKKγ/NEMO (#I5032), anti-IKKγ/NEMO (#WH0008517M1), anti-β-actin (Sigma-Aldrich, Germany), and anti-UK (Invitrogen, Germany). Recombinant human TNFα was purchased from Miltenyi (Bergisch Gladbach, Germany), and the DNA-PK inhibitor NU7026 was purchased from Calbiochem (La Jolla, CA, USA).

The following antibodies were used: anti-GSK-3β (#9832), anti-K48-linked ubiquitin (#8081), NF-κB Pathway Sampler Kit (#9936), anti-TNF-R1 (#C25C1) and anti-RIP (#D94C12) (New England BioLabs, Frankfurt/Main, Germany), anti-Ubiquitin from DAKO (Hamburg, Germany), anti-pSer-31-NEMO, anti-TRAF6, anti-RIP1 and anti-β-actin (Abcam, Cambridge, UK); anti-p47 (Abnova, Heidelberg, Germany); phospho-specific anti-pSer-8-NEMO and anti-pSer-17-NEMO (Eurogentec, Köln, Germany); anti-FLAG M2 mouse (Sigma-Aldrich, Taufkirchen, Germany); anti-pSer-43-NEMO (Abgent, Heidelberg, Germany); anti-Tubulin and non-immune IgGs (Invitrogen, Darmstadt, Germany). Recombinant human TNFα was from Miltenyi (Bergisch Gladbach, Germany), GSK-3 inhibitors SB216730 and AR-A014418 from Calbiochem (La Jolla, CA, USA).