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. Author manuscript; available in PMC: 2020 Dec 1.
Published in final edited form as: Mol Cancer Res. 2020 Feb 21;18(6):913–925. doi: 10.1158/1541-7786.MCR-19-0726

Figure 3. Identification of Rho GTPase GEFs necessary for migration and Cdc42 activation using an siRNA based screen.

Figure 3.

A. MiaPaCa2 cells were transiently transfected with individual siRNA from a GEF siRNA library, and migration towards DRG by Boyden chamber assay was measured. The GEFs required for migration (below the red line) were selected. B. Cell proliferation was assessed following siRNA transfection of selected GEFs to exclude the GEFs required for proliferation. Resulting GEF candidates (above the red line) were selected. ECT2 was excluded because silencing resulted in a multinucleated cell phenotype. C. Each GEF candidate underwent siRNA tranfection followed by quantification of GTP-Cdc42 (active Cdc42) by G-LISA following stimulation with GDNF. The final 6 GEFs (below the red line) meet the criteria of: (1) driving cancer cell migration towards DRG, (2) not required for cell proliferation, and (3) activating Cdc42.