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. 2019 Aug 27;10(20):4777–4792. doi: 10.7150/jca.26740

Figure 4.

Figure 4

Figure 4

In HCCLM3 cells, Fucoidan-Sargassum inhibits the maturation stages of invadopodia formation, deactivates integrin αVβ3 and transcription factor E2F1, induces endoplasmic reticulum stress, and enhances cell-ECM adhesion. (A-C) Western blotting analysis of the integrin receptors αVβ3, the transcription factor E2F1, the key proteins of endoplasmic reticulum stress GRP78, IRE1, and SPARC, and ECM collagen-related integrin α1β1 and cell adhesion proteins E-Cadherin and Caveolin-1, after the cells were treated with Fucoidan-Sargassum (0, 10, 20, 30 mg/mL) for 48 h. (D) Confocal images of fixed control cells or Fucoidan-Sargassum-treated cells immunostained for actin (2nd panel), cortactin (middle panel), and integrin αV (4th panel). The arrowheads indicate active invadopodia in the control cells and cytoskeletal changes in the treated cells. (E) Confocal images of fixed control cells or Fucoidan-Sargassum-treated cells immunostained for actin (2nd panel), and SPARC (3rd panel). Note that the localization of SPARC in the control and treated cells is identified by arrowheads. Scale bars: 7.5 µm. (F) Western blotting analysis of ITGαVβ3/Src/E2F1 signaling pathway after the cells were transfected with shITGαV and shITGβ3, and treated with Fucoidan-Sargassum (0, 10, 20, 30 mg/mL) for 48 h.