Negative Effects of SIGIRR on TRAF6 Ubiquitination in Acute Lung Injury In Vitro

Figure 1

Analysis by pull-down and coimmunoprecipitation experiments of the interaction between SIGIRR and TRAF6 and resultant (auto) ubiquitinylation of TRAF6. (a) GST pull-down assay was performed using purified GST-fusion SIGIRR and His-tagged TRAF6 proteins. (b) A lysate from HEK293 cells cotransfected with Flag-TRAF6 or HA-SIGIRR was subjected to immunoprecipitation with anti-HA or anti-Flag antibody, respectively. (c) A lysate from HEK293 cells cotransfected with SIGIRR, Flag-TRAF6, or HA-ubiquitin, respectively, was subjected to immunoprecipitation with anti-Flag antibody. (d) HEK293 cells were cultured and cotransfected with the above regents. (e) Colocalization between SIGIRR and TRAF6 in HEK293 cells was also evaluated by immunofluorescence confocal microscopy.

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