Rasd1 Modulates the Coactivator Function of NonO Figure 3. GTP hydrolysis activity of Rasd1 is required to cooperate with NonO to suppress CREB’s activity. (A) NonO localises primarily in the nucleus of HEK293T cells transiently transfected with pNonO-V5 (2 mg) (Figure A2). Rasd1 is distributed throughout the cell in the event of individual transfection of pHis-Rasd1 (2 mg) (Figure A5). A considerable increase in the amount of Rasd1 was observed to be present in the nucleus upon the event of co-transfection with pGST-NonO (compare Figures A5 with A8). The sub-cellular location of NonO was unaffected in cells co- transfected with pHis-Rasd1 (compare Figures A2 with A9). Rasd1 mutants, A178V, G81A, T38N and Del-NLS, display similar cellular distribution to PLoS ONE | www.plosone.org 7 September 2011 | Volume 6 | Issue 9 | e24401