FIGURE 5:

Importance of the C1 domain for the RalA binding. (A) Direct binding of FLAG-RalA to GST-RD, but not to GST-ΔC1. Pull-down assay using FLAG-RalA and GST-RD or GST-ΔC1 was performed as described in Materials and Methods. (B) Colocalization of FLAG-RalA with RD-GFP. NHEK cells expressing FLAG-RalA were subsequently infected with viruses coding RD-GFP (RD), ΔC1-GFP (ΔC1), or GFP alone. The cells were fixed 24 h after the infection, followed by immunocytochemistry using anti–FLAG antibody and Alexa 594–conjugated secondary antibody. Both red and green fluorescences were observed under confocal microscopy.