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Fig. 6. Cellular localisation of GFP-4.1B fusion protein. To investigate the localisation of 4.1B, cells were transfected with GFP-4.1B fusion construct and, 24 hours later, fixed and stained with Rhodamine-phalloidin. (A-D) Sarcoma cells K2. (E-L) HeLa cells. (A) GFP-4.1B is expressed in the cytoplasm and is enriched at the plasma membrane. (B,C) There is also some colocalisation with F-actin especially at the plasma membrane compartment enriched with GFP-4.1B. The brighter area of GFP-4.1B between the two cells in A is probably a result of the two cells overlapping, suggested by the phase-contrast image (D). (E,F) Localisation of 4.1B in HeLa cells is similar, with cytoplasmic expression and some colocalisation with F-actin (G), especially in peripheral areas of ruffling and protrusive activity, as seen in the enlarged region (H; boxed region in G). There is also an enrichment of GFP-4.1B at the membrane (I-K). This is clearer still in the close-up (L) of the boxed region in (I). All images are single confocal images and representative of three independent experiments. Bar, 20 µm.
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