Figure 6

Distribution of phosphomimetic mutants of FIP3 and their effect on transferrin uptake in interphase cells. HeLa cells on glass coverslips were transiently transfected with GFP-FIP3 or the indicated mutants as described in Methods. 48 h later, cells were incubated for two hours in serum free D-MEM, transferred to media supplemented with 1 μg/ml human transferrin conjugated to Texas Red and incubated at 37°C for 30 minutes. Cells were fixed and the distribution of GFP-FIP3 (pseudo-coloured green) and Tf (pseudo-coloured red) analysed by confocal microscopy. Merged images were generated using the Zeiss confocal software. Shown are representative images for each mutant. Over 50 cells were analysed for each mutant at each condition, and each experiment was performed in triplicate.