Fig. 7. Colocalization of clathrin and EGFRh in cells expressing the dominant-negative Rab5-CAAX mutant. CFP-Rab5-CAAX and CFP-Rab5(S34N)-CAAX were transiently co-transfected with YFP-CL_A in PAE/EGFR cells. The cells were incubated with EGF-Rh as in Fig. 6. EGF-Rh, YFP and CFP were detected using Cy3, YFP and CFP filter channels. 'Yellow' signifies colocalization of YFP and rhodamine fluorescence, whereas 'cyan' signifies colocalization of CFP and rhodamine fluorescence. Insets show an enlargement of the outlined regions. Additional insets below the YFP image show the merge image of EGF-Rh and ß2-YFP, in which the EGF-Rh image (red) was right-shifted 4 pixels (250 nm) to allow better visualization of colocalization of red and green dots. The arrows show an example of EGF-Rh-containing endosomes in a cell that does not express CFP-Rab5(S34N)-CAAX.

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