Fig. 3. Localisation of WEF-binding pocket MO25 mutants in cells. HeLa cells were transfected with the construct encoding wild-type or indicated mutants of Myc-MO25 in the absence or presence of GFP-LKB1 and Flag-STRAD. Twenty-four hours post-transfection, the cells were fixed in 4% (v/v) paraformaldehyde and immunostained with the anti-MO25 antibody to detect MO25 (TR anti-sheep secondary antibody, red channel) and anti-Flag antibody to detect STRAD (Cy5 anti-mouse secondary antibody, blue channel). GFP-LKB1 localization was visualized directly through the GFP fluorescence (green channel). The cells were imaged using a Zeiss LSM 510 META confocal microscope. The cells shown are representative images obtained in three separate experiments.