Figure 2: Infected cells retain functionality.
From: Highly efficient baculovirus-mediated multigene delivery in primary cells
(a) COS7 cells were infected with MultiPrime baculoviruses expressing the indicated fluorescently-tagged RAB GTPases. Cells were stimulated for 3 h with Cy5-labelled EGF. As expected, EGF was found in RAB7A vesicles (arrows) and RAB5A vesicles (arrowheads) but not in RAB11 vesicles. A quantitative time-resolved analysis is provided (Supplementary Movie 1). (b) PAE cells stably expressing VEGFR2 and neuropilin-1 were infected with baculoviruses expressing the indicated fluorescently-tagged RAB GTPases. Cells were then stimulated with Nt647-labelled VEGF. VEGF can be found in RAB5 and Rab7 vesicles (arrows). (c) Tube formation: HUVEC were infected with a baculovirus expressing mTFP1-actin, EYFP-tubulin and Mito-dsRed (only red channel is shown). DAPI was used to counterstain nuclei of all cells. Approximately 30% of cells were infected. Infected and uninfected cells contribute to tubes. (d) Migration: HUVEC were infected with a baculovirus expressing mTFP1-actin, EYFP-tubulin and Mito-dsRed (only red channel is shown). Approximately 30% of cells were infected. Infected and uninfected cells migrate at same rates. Scale bar, 20 μm (a,b); 500 μm (c,d).
