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. 2012 May 8;7(5):e36601. doi: 10.1371/journal.pone.0036601

Figure 5. The ability of K13 to activate NF-κB pathway does not require linear polyubiquitination of NEMO. A–B.

Figure 5

MEF NEMO−/Y cells were stably transduced with wild-type NEMO and two NEMO mutants defective in linear ubiquitination of NEMO (NEMO-Y293A/Y301A/K302A and NEMO-F305A) and their expression was confirmed by immunoblotting using NEMO antibody. These cells were further transduced with either an empty retroviral vector or a retroviral vector expressing FLAG-tagged K13 and their expression was determined by immunoblotting with FLAG antibody. The blot was re-probed with a tubulin antibody to show equal loading of protein. C. Upon expression of K13, these cells were able to activate NF-κB p65 DNA binding as determined by ELISA. The values shown are averages (mean±SE) of one representative experiment out of three in which the level of p65 DNA binding was measured in triplicate. Asterisks (*) indicate significance at levels of p≤0.05.