Table 2.
Import of T-DNA complex into mammalian nuclei occurs via the classical NLS-dependent import pathway
| Import substrate | Inhibitor | Lacking factor | Import |
|---|---|---|---|
| ssDNA–VirD2–VirE2 | — | — | + |
| ssDNA–VirD2–VirE2 | — | Rch1 | − |
| ssDNA–VirD2–VirE2 | — | Ran | − |
| ssDNA–VirD2–VirE2 | — | GTP/ATP | − |
| ssDNA–VirD2–VirE2 | WGA | — | − |
| ssDNA–VirD2–VirE2 | GTPγS | — | − |
The import assay of fluorescently labeled T-DNA complex (ssDNA–VirD2–VirE2) into mammalian nuclei was done in vitro using permeabilized HeLa cells. Each experiment was repeated at least three times, and for each sample at least 100 nuclei were scored. +, 99–100% of nuclei showed red fluorescence signal; −, 0–1% of nuclei showed red fluorescence signal. GTPγS, guanosine 5′−[γ−thio]triphosphate. WGA, wheat germ agglutinin.