Figure 3. TRAF6-ECSIT signaling regulates generation of mitochondrial and cellular ROS, which requires TRAF6 E3-ubiquitin ligase activity.
a-c, WT or ECSIT +/- BMM were transduced with shRNAs and left untreated or stimulated with LPS. Untreated BMM were lysed and extracts blotted for TRAF6 and ECSIT (a). Cells were stained with MitoSOX (b) or CM-H2DCFDA (c) and analyzed by FACS. d-f, WT or TRAF6 null BMM were left untreated or transduced with TRAF6 expressing retroviruses, then stimulated with for the indicated times. Unstimulated BMM were lysed and extracts blotted for TRAF6 expression (d). Cells were stained with MitoSOX (e) and CM-H2DCFDA (f) and analyzed by FACS. Error bars represent s.d. of the mean from triplicate samples.