Fig. 4. The presence of transmembrane domains is sufficient to direct trafficking of L6 to late endocytic organelles. (A) Schematic diagram shows the constructs used for transfection of MCF-7 cells. (B) MCF-7 cells were transfected with the plasmid encoding the L6H-HA, L6-Ag-HA and L6-L6H chimeras, and 48 hours later the cells were processed for double-immunofluorescence staining using specific Abs as described in the legend to Fig. 2. Co-distribution of the tagged proteins with Lamp2 was analysed using anti-HA mAb F7 (IgG2a) and anti-Lamp2 mAb H4B4 (IgG1). Staining was visualised using Alexa-Fluor-594-conjugated goat anti-mouse IgG2a Ab (red) and Alexa-Fluor-488-conjugated goat anti-mouse IgG1 (green). Scale bar: 20 µm.