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. 2011 Dec 19;109(1):191–196. doi: 10.1073/pnas.1105176108

Fig. 2.

Fig. 2.

Miz1 interferes with the interaction between TRAF2 and Ubc13. (A) WT fibroblasts were transfected with expression vectors encoding HA-TRAF2, HA-ΔRING-TRAF2 mutant (ΔR), or HA-ΔTRAF-C-TRAF2 mutant (ΔC) (5 μg each), along with Xpress-Miz1 or Xpress-Ubc13 (5 μg each). The interaction between various HA-TRAF2 and Miz1 or Ubc13 was analyzed. (B) HEK293 cells were transfected with expression vectors encoding Xpress-Miz1 or Xpress-Ubc13, along with WT M2-TRAF2 or M2-TRAF2 RING domain mutant (23). The interaction between Xpress-Miz1 or Xpress-Ubc13 with WT or the RING domain mutant of M2-TRAF2 was analyzed. (C) WT fibroblasts were transfected with expression vectors encoding M2-TRAF2, along with HA-Ubc13 (5 μg each), with or without various amounts of Xpress-Miz1 (3, 4, and 6 μg), as indicated. The interaction between M2-TRAF2 and HA-Ubc13 or Xpress-Miz1, as well as ubiquitination of M2-TRAF2, in control or TNFα-stimulated cells was analyzed. (D) The association between TRAF2 and Miz1 or Ubc13 in control or TNFα-treated WT and Miz1-null MEFs were analyzed. (E) The effect of Ubc13 knockdown or double knockdown of Ubc13 and Miz1 on TNF-α–induced JNK phosphorylation was analyzed.