In Vivo Identification of Regulators of Cell Invasion Across Basement Membranes

David Q. Matus, Xiao-Yan Li, Sarah Durbin, Daniel Agarwal, Qiuyi Chi, Stephen J.Weiss, David R. Sherwood*

*To whom correspondence should be addressed. E-mail: david.sherwood{at}duke.edu

This PDF file includes:

• Fig. S1. Putative null alleles of mep-1, hda-1, cct-6, C48A7.2, and hbl-1 show AC invasion defects.
• Fig. S2. LAM-1::GFP (laminin) is intact following RNAi targeting the cct complex and 11 other pro-invasive genes at the P6.p 4-cell stage of VPC division.
• Fig. S3. Transgene reporter localization of newly identified pro-invasive genes at the P6.p 2-cell stage of VPC division.
• Fig. S4. Transgene reporter localization of newly identified pro-invasive genes at the P6.p 4-cell stage of VPC division.
• Fig. S5. hbl-1 functions in VPC specification to promote AC invasion.
• Fig. S6. The AC is correctly specified, as shown by lin-3>GFP, following RNAi depletion of the newly identified regulators of AC invasion.
• Fig. S7. 1° VPC specification following RNAi depletion of newly identified AC invasion genes.
• Fig. S8. Identification of regulators of invasive membrane formation.
• Fig. S9. Regulators of invasive membrane formation function independently of netrin receptor localization.
• Fig. S10. Regulators of invasive membrane formation function independently of integrin receptor localization.
• Fig. S11. Identification of regulators of the fos-1a pathway.
• Fig. S12. Identification of genes that act within or parallel to the fos-1a pathway.
• Fig. S13. fos-1a RNAi depletion identifies lit-1 as a new member of the fos-1a pathway.
• Fig. S14. lit-1 functions downstream of fos-1a and egl-43L during AC invasion.
• Fig. S15. CCT5 and NLK siRNA-mediated knockdown specifically affect BM transmigration activity.
• Table S1. 539 Pvl and Egl genes targeted by RNAi for AC invasion defects.
• Table S2. Identification of 99 regulators of AC invasion.
• Table S3. RNAi depletion of 99 genes results in an AC invasion defect.
• Table S4. Timing and degree of AC invasion into the vulval epithelium: mutant analysis and off-target RNAi controls.
• Table S5. Timing and degree of AC invasion into the vulval epithelium: uterine-specific RNAi.
• Table S6. Extrachromosomal array and integrated strain generation.
• Table S7. Primer sequences and templates used for PCR fusions and restriction enzyme cloning.

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Other Supplementary Material for this manuscript includes the following:

• Table S1. 539 Pvl and Egl genes targeted by RNAi for AC invasion defects (Excel file).

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Technical Details

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© 2010 American Association for the Advancement of Science