Figure 5.

Development of a NAEK-containing lentiviral vector as a tool for single-virus tracking. (A) Characterization of the visibility of NAEK-containing lentiviral vectors at K242 upon treatment with DIBO-Alexa 488. The wild-type vectors (WT) treated by DIBO-Alexa 488 and NAEK-containing lentiviral vector without treating by DIBO-Alexa 488 act as the negative control. (B) Real-time monitoring of the dynamic process of entry and transport of Alexa 488-conjugated lentiviral vector. HeLa cells were stained with Hoechst 33342 and DiI, respectively, to label host nucleic acids and the plasma membrane. After incubation of HeLa cells with lentiviral vector at 4°C for 30 min to synchronize binding, the infected HeLa cells were placed at 37°C in a CO₂ online culture system and tracked using a spinning-disk confocal microscope platform. The Alexa 488 was detected at 488 nm, and DiI was detected at 555 nm. A series of images were selected from 1 to 18 min that reflect the dynamic process of endocytosis of lentiviral vectors.