Supplementary Materials

Supplementary Materials for:

Prostaglandin E2–EP2–NF-κB signaling in macrophages as a potential therapeutic target for intracranial aneurysms

Tomohiro Aoki, Juhana Frȍsen, Miyuki Fukuda, Kana Bando, Go Shioi, Keiichi Tsuji, Eliisa Ollikainen, Kazuhiko Nozaki, Johanna Laakkonen, Shuh Narumiya*

*Corresponding author. Email: snaru{at}mfour.med.kyoto-u.ac.jp

This PDF file includes:

  • Fig. S1. Scoring of COX-2 and EP2 abundance in human intracranial aneurysm lesions.
  • Fig. S2. Immunostaining for EP2, CD31, and/or α–smooth muscle actin in human intracranial aneurysm lesions.
  • Fig. S3. Verification of the NF-κB reporter construct.
  • Fig. S4. NF-κB activation in intracranial arterial wall during intracranial aneurysm development assessed in NF-κB reporter mice.
  • Fig. S5. Generation of the Ptger2flox/flox mouse line.
  • Fig. S6. Effect of macrophage-specific deletion of Ptger2 on MCP-1 and COX-2 immunofluorescence in intracranial aneurysm lesions.
  • Fig. S7. Generation of the IκB mutant transgenic mouse line.
  • Fig. S8. Cre-dependent expression of the IκB mutant and suppression of NF-κB activation.
  • Fig. S9. Generation of transgenic mouse lines expressing a macrophage-specific IκB mutant–mCherry fusion protein.
  • Fig. S10. Generation of transgenic mouse lines expressing IκB mutant–mCherry fusion protein in an endothelial cell–specific fashion.
  • Fig. S11. Effects of macrophage- and endothelial cell–specific expression of the IκB mutant protein on systemic blood pressure and macrophage recruitment after intracranial aneurysm induction.
  • Fig. S12. Generation of the HEK293-EP2 cell line.
  • Fig. S13. NF-κB activation in response to EP2 signaling in HEK293-EP2 cells.
  • Fig. S14. Involvement of NF-κB activation in amplifying the effect of EP2 on TNF-α–induced gene expression.
  • Fig. S15. Analysis of the Ptgs2 gene promoter for κB sites responsive to EP2 and TNF-α signaling.
  • Fig. S16. Activation of Sp1 by TNF-α signaling in HEK293-EP2 cells.
  • Fig. S17. EP2-induced HuR translocation.
  • Fig. S18. Activation of the MAPK p38 by EP2 signaling in HEK293-EP2 cells.
  • Fig. S19. Effects of PF-04418948 on systemic blood pressure and thinning of media in intracranial aneurysm walls.
  • Table S1. Gene expression analysis of rat intracranial aneurysm lesions.

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Other Supplementary Material for this manuscript includes the following:

  • Data File S1 (Microsoft Excel format). Full gene list from microarray analysis of rat intracranial aneurysm lesions.

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Citation: T. Aoki, J. Frȍsen, M. Fukuda, K. Bando, G. Shioi, K. Tsuji, E. Ollikainen, K. Nozaki, J. Laakkonen, S. Narumiya, Prostaglandin E2–EP2–NF-κB signaling in macrophages as a potential therapeutic target for intracranial aneurysms. Sci. Signal. 10, eaah6037 (2017).

© 2017 American Association for the Advancement of Science