Novel p47^phox-Related Organizers Regulate Localized NADPH Oxidase 1 (Nox1) Activity

Davide Gianni, Begoña Diaz, Nicolas Taulet, Bruce Fowler, Sara A. Courtneidge,* Gary M. Bokoch*

*To whom correspondence should be addressed. E-mail: bokoch{at}scripps.edu (G.M.B.) and courtneidge{at}burnham.org (S.A.C.)

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• Fig. S1. Myc-tagged transfected organizer subunits are expressed in comparable amounts in reconstituted HEK293 cells.
• Fig. S2. DLD1 cells only express the Nox1 member of the NADPH oxidase family, the NoxA1 activator, and Tks4 (and small amounts of Tks5) as members of the p47^phox organizer superfamily.
• Fig. S3. The presence of Tks5 single SH3 domain inactivating mutations (M1 to M5), or the TksM1M2 double mutation did not affect the ability of Tks5 to support Nox1-dependent ROS generation and to bind NoxA1.
• Fig. S4. Tks4-specific siRNA efficiently knocks down Tks4 protein at 72 and 96 hours and significantly reduces the ability of DLD1 cells to degrade ECM.
• Fig. S5. Nox1-specific siRNAs efficiently block ROS generation, invadopodia formation, and ECM degradation in DLD1 cells and this effect can be rescued by overexpression of murine Nox1 (mNox1).
• Fig. S6. In DLD1 cells, Nox1 and phalloidin- and Nox1 and cortactin-positive structures are ventral organelles that have the characteristics of invadopodia.
• Fig. S7. NoxO1-GFP does not localize to invadopodia in DLD1 cells and invadopodia formation in NoxO1-transfected cells is greatly reduced.

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Citation: D. Gianni, B. Diaz, N. Taulet, B. Fowler, S. A. Courtneidge, G. M. Bokoch, Novel p47^phox-related organizers regulate localized NADPH oxidase 1 (Nox1) activity. Sci. Signal. 2, ra54 (2009).

© 2009 American Association for the Advancement of Science