Fig. 6. Subcellular localisation of MARK3. (A) Wild-type (wt) or indicated mutant forms of GST-MARK3 (see Fig. 5) were expressed in HEK 293 cells, which were fractionated into a cytosolic and a membrane fraction. Subcellular fractions, including solubilised portions of the homogenates, were subsequently analysed by western blot with regards to MARK3 (GST), MARK3 T-loop phosphorylation (p-T211), the cytosolic marker GAPDH (GAPDH) and the plasma membrane marker Na-K-ATPase (Na-K-ATPase). The results are representative of three experiments. (B) Wild-type (wt) or indicated mutant forms of GFP-MARK3 (see Fig. 5) were expressed in HEK 293 cells, which were fixed in 3% paraformaldehyde 24 hours post transfection. The GFP-fluorescence was analysed by confocal fluorescence microscopy. The cells shown are representative of images obtained in two separate experiments. Bars, 10 µm.