Supplementary Materials

Supplementary Materials for:

The Secreted Protein ANGPTL2 Promotes Metastasis of Osteosarcoma Cells Through Integrin α5β1, p38 MAPK, and Matrix Metalloproteinases

Haruki Odagiri, Tsuyoshi Kadomatsu, Motoyoshi Endo, Tetsuro Masuda, Masaki Suimye Morioka, Shigetomo Fukuhara, Takeshi Miyamoto, Eisuke Kobayashi, Keishi Miyata, Jun Aoi, Haruki Horiguchi, Naotaka Nishimura, Kazutoyo Terada, Toshitake Yakushiji, Ichiro Manabe, Naoki Mochizuki, Hiroshi Mizuta, Yuichi Oike*

*Corresponding author. E-mail: oike@gpo.kumamoto-u.ac.jp

This PDF file includes:

  • Fig. S1. Expression of genes encoding DNA demethylation–associated enzymes in tumor cells maintained in hypoxic and serum-starved conditions.
  • Fig. S2. ANGPTL2 expression and promoter methylation status in human breast cancer cell lines.
  • Fig. S3. ANGPTL2 expression and promoter methylation status in human pancreatic cancer cell lines.
  • Fig. S4. ANGPTL2 expression in OS cell lines correlates with lung metastasis in an orthotopic xenograft model.
  • Fig. S5. ANGPTL2 knockdown.
  • Fig. S6. Expression of MMPs and phosphorylation of p38 MAPK in ANGPTL2-deficient cells.
  • Fig. S7. Effect of ANGPTL2 overexpression on OS cell proliferation and tumor volume.
  • Fig. S8. ANGPTL2 promotes metastasis in an orthotopic xenograft model.
  • Fig. S9. Effect of a pan-MMP inhibitor on the invasive activity of tumor cells.
  • Fig. S10. ANGPTL2 activates p38 MAPK signaling through integrin α5β1.
  • Fig. S11. Effect of inhibitors of IKKβ and p38 MAPK on MMP activity in ANGPTL2-overexpressing 143B cells.
  • Fig. S12. Detection of LILRB2 on 143B cells.
  • Fig. S13. ANGPTL2 knockdown in tumor cells decreases tumor angiogenesis and tumor cell intravasation.
  • Fig. S14. ANGPTL2 does not disrupt endothelial monolayers or promote tumor cell extravasation.
  • Fig. S15. ANGPTL2 abundance in primary tumor tissue correlates with lung metastasis and MMP-9 abundance.
  • Fig. S16. ANGPTL2 mRNA and protein abundance in cell lines overexpressing full-length or fragment constructs.
  • Fig. S17. ANGPTL2 is not cleaved by pro-protein convertases.
  • Fig. S18. ANGPTL2 and TLL1 expression in OS cell lines.
  • Fig. S19. ANGPTL2 and TLL1 expression in clinical samples of OS.
  • Table S1. Classification of OS patients based on primary tumor ANGPTL2 and MMP-9 expression.
  • Table S2. Primer pairs used to construct expression vectors encoding CCD or FLD ANGPTL2 fragments or mutant ANGPTL2.
  • Table S3. Primer pairs used for real-time PCR analysis.
  • Table S4. Primer pairs used for DNA methylation analysis and genomic sequencing.
  • Reference (64)

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Citation: H. Odagiri, T. Kadomatsu, M. Endo, T. Masuda, M. S. Morioka, S. Fukuhara, T. Miyamoto, E. Kobayashi, K. Miyata, J. Aoi, H. Horiguchi, N. Nishimura, K. Terada, T. Yakushiji, I. Manabe, N. Mochizuki, H. Mizuta, Y. Oike, The Secreted Protein ANGPTL2 Promotes Metastasis of Osteosarcoma Cells Through Integrin α5β1, p38 MAPK, and Matrix Metalloproteinases.. Sci. Signal. 7, ra7 (2014).

© 2014 American Association for the Advancement of Science