Figure 6. GefE activates RasD.

(A) Comparison of the levels of activated RasD-GTP and total RasD by Western blot in vegetative or 12 hr starved cells grown in the presence or absence of glucose. (B) Comparison of activated RasD-GTP and total RasD levels by Western blot in wild type Ax3, gefE⁻ or rasD⁻ cells. (C) GFP-labelled (*) Ax3 wild type cells mixed at 10:90 ratio with unlabelled Ax3 or rasD⁻ mutant cells. Closed arrows show enrichment of wild type cells in pstO and pstB populations. Open arrows show reciprocal enrichment of rasD⁻ cells. (D) RasD overexpression results in GefE dependent bias towards the pstO and pstB cell fates. 20% cells constitutively expressing RFP, RFP-RasD or RFP-RasD(G12T) were mixed with 80% unlabelled parental cells. When wild type Ax3 cells overexpress RFP-RasD or RFP-RasD(G12T) they become enriched in the pstO and pstB populations (arrows). Only gefE⁻ cells that express constitutively activated RasD(G12T) are enriched in pstO and pstB populations (arrows).

DOI: http://dx.doi.org/10.7554/eLife.01067.013

Figure 6—figure supplement 1. rasD⁻ mutant cells avoid pstO and pstB fates in chimera with wild type cells.

10% Ax3 labelled wild type were mixed with 90% unlabelled rasD⁻ mutant cells. The top row shows the control mix and is comparable to the image directly beneath. Closed arrows indicate enrichment of label, open arrows an absence. The AP axis of all slugs is aligned from right-left.

DOI: http://dx.doi.org/10.7554/eLife.01067.014