Supplementary Materials

Supplementary Materials for:

Depletion of the 26S Proteasome Adaptor Ecm29 Increases Toll-Like Receptor 3 Signaling

Carlos Gorbea,* Martin Rechsteiner, Jesús G. Vallejo, Neil E. Bowles*

*Corresponding author. E-mail: neil.bowles@hsc.utah.edu (N.E.B.); gorbea@biochem.utah.edu (C.G.)

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  • Fig. S1. siRNA duplexes targeting sequences within the 3′ untranslated region of Ecm29 mRNA do not reduce Ecm29 abundance.
  • Fig. S2. Analysis of the amounts of TLR3, p62, and LC3β in cells transfected with siRNAs that do not knock down Ecm29.
  • Fig. S3. Dose-dependent stimulation of control HeLa cells with poly(I:C).
  • Fig. S4. Distribution of TLR3, LC3β, p62, and Ecm proteasomes in control and Ecm29-depleted cells.
  • Fig. S5. Knockdown of Ecm29 decreases the vesicular localization of TRIF and increases its association with a juxtanuclear membrane compartment.
  • Fig. S6. Knockdown of Ecm29 decreases the vesicular localization of TRAF3 and increases its association with a juxtanuclear membrane compartment.
  • Fig. S7. Knockdown of Ecm29 decreases the vesicular localization of TBK1 and increases its association with a juxtanuclear membrane compartment.
  • Fig. S8. Knockdown of Ecm29 decreases the vesicular localization of pIKKε and increases its association with a juxtanuclear membrane compartment.
  • Fig. S9. Distribution of TRIF, TRAF3, TBK1, and pIKKε in control and Ecm29-depleted cells.
  • Fig. S10. Accumulation of pTBK1 in an Arf6-containing compartment in Ecm29-depleted cells.
  • Fig. S11. Colocalization of TBK1 with Rab11 and Rab7 in poly(I:C)-stimulated TLR3-HA HeLa cells transfected with control siRNA.
  • Fig. S12. Ecm29 localizes to Rab7-, Rab11-, and Arf6-containing vesicles upon stimulation of TLR3.
  • Fig. S13. GFP-Ecm29(882–1319) does not inhibit the degradation of polyubiquitin protein conjugates.
  • Fig. S14. Displacement of Ecm proteasomes from endosomes and autophagic vesicles increases the extent of TLR3 signaling.
  • Fig. S15. GFP-Ecm29(882–1319) prevents the localization of the 26S proteasome to vesicles containing pIRF3.
  • Fig. S16. GFP-Ecm29(882–1319) prevents the localization of the 26S proteasome to vesicles containing pTBK1.
  • Fig. S17. GFP-Ecm29(882–1319) prevents the localization of the 26S proteasome to vesicles containing pIKKε.

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Citation: C. Gorbea, M. Rechsteiner, J. G. Vallejo, N. E. Bowles, Depletion of the 26S Proteasome Adaptor Ecm29 Increases Toll-Like Receptor 3 Signaling. Sci. Signal. 6, ra86 (2013).

© 2013 American Association for the Advancement of Science