Fig 8.
The Asp14 C-terminal region binds mammalian host cells. RF/6A cells were incubated with GST-Asp14, GST-Asp141–64, GST-Asp1465–124, GST-Asp141–100, GST-Asp141–112, GST-APH_1387112–579 (negative control; does not associate with mammalian cell membranes), or GST alone for 60 min, followed by removal of unbound protein. (A) Confocal microscopy analyses of GST fusion proteins bound to RF/6A cells. The host cells were fixed and successively incubated with anti-GST antibody and anti-mouse IgG conjugated to Alexa Fluor 594. Gel mounting medium containing DAPI was added. Representative merged fluorescent images are shown. Results are representative of two to four independent experiments. (B) Flow cytometric analysis of GST fusion protein binding to RF/6A cells. The host cells were successively incubated with GST antibody and Alexa Fluor 488-conjugated anti-mouse IgG and then analyzed by flow cytometry.