PKCε Regulation of an α₅ Integrin–ZO-1 Complex Controls Lamellae Formation in Migrating Cancer Cells

Saara Tuomi, Anja Mai, Jonna Nevo, Jukka O. Laine, Vesa Vilkki, Tiina J. Öhman, Carl G. Gahmberg, Peter J. Parker, Johanna Ivaska*

*To whom correspondence should be addressed. E-mail: Johanna.ivaska{at}vtt.fi

This PDF file includes:

• Materials and Methods
• Fig. S1. α₅ integrin is increased on the cell surface and contributes to migration of NCI-H460 cells.
• Fig. S2. α₅ integrin, but not α₂ integrin, is recruited to the lamellae with ZO-1 upon wounding.
• Fig. S3. Verification of siRNA efficacy.
• Fig. S4. α₅ integrin and ZO-1 colocalize in the lamellae of breast cancer cells; silencing of ZO-1 influences focal adhesion morphology.
• Fig. S5. ZO-1 knock-down increases Rac activity.
• Fig. S6. α₅WT- and α₅PPAA-expressing cells adhere equally well to fibronectin but differ in lamellae formation.
• Fig. S7. α₅ integrin and ZO-1 form a complex in a fraction of lung cancers.
• Fig. S8. ZO-1 phosphorylation is increased in subconfluent cells; PKC inhibition impairs recruitment of α₅ integrin and ZO-1 to lamellae; α₅ mediates adhesion to fibronectin.
• Fig. S9. Loss of PKCε does not influence internalization of α₅ integrin from the cell surface.
• Fig. S10. PKCε regulates the subcellular localization of ZO-1 through the putative PKC site Ser¹⁶⁸.

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Technical Details

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Other Supplementary Materials for this manuscript include the following:

Movies S1 through S5 and movie descriptions

• Descriptions of movies S1 to S5. (Adobe Acrobat PDF; file size 119 KB)
• Movie S1. (filename: Movie_S1.mov; file size 896 KB; QuickTime Movie)
• Movie S2. (filename: Movie_S2.mov; file size 5.98 MB; QuickTime Movie)
• Movie S3. (filename: Movie_S3.mov; file size 5.67 MB; QuickTime Movie)
• Movie S4. (filename: Movie_S4.mov; file size 876 KB; QuickTime Movie)
• Movie S5. (filename: Movie_S5.mov; file size 1.11 MB; QuickTime Movie)

Citation: S. Tuomi, A. Mai, J. Nevo, J. O. Laine, V. Vilkki, T. J. Öhman, C. G. Gahmberg, P. J. Parker, J. Ivaska, PKCε regulation of an α₅ integrin–ZO-1 complex controls lamellae formation in migrating cancer cells. Sci. Signal. 2, ra32 (2009).

© 2009 American Association for the Advancement of Science