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Fig. 4. Extension of SNX5-positive tubules is dependent on microtubules. (A) HEK-GFP-SNX5 cell monolayers were fixed in 0.9% PFA and analysed by indirect immunofluorescence using a monoclonal antibody against ß-tubulin followed by Cy3-conjugated goat anti-mouse IgG. (B) HEK-GFP-SNX5 cell monolayers were serum starved overnight before being exposed to the microtubule-destabilising agent nocodazole (10 µM) for 30 minutes, and then to EGF as in Fig. 3. Time-lapse movies were recorded for a total period of 15 minutes with a 10-second interval between image capture. The total capture period was subsequently extended to 90 minutes with little apparent loss of fluorescent signal of GFP-SNX5 from the macropinosome. Asterisks indicate the lumen of an individual macropinosome. Bars, 5 µm (A); 10 µm (B).
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