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. 2009 Jun 5;5(6):e1000465. doi: 10.1371/journal.ppat.1000465

Figure 4. Decreased co-localization of Py with the late endosome and lysosome in GD1a-supplemented cells.

Figure 4

(A) NIH 3T3 cells were incubated with purified GM1 or GD1a, washed, infected with Py, and the extent of infection was assessed as in Figure 1B. Results were normalized to non-supplemented cells (control cells). In the control cells, 43/984 cells expressed large T antigen. (B) Untreated (control) or GD1a-supplemented NIH 3T3 cells were incubated with Py at 4°C to allow viral binding and then treated with proteinase K where indicated (top panel) or incubated at 37°C for 1 hr before proteinase K treatment to determine viral entry (bottom panel). (C, D) The extent of co-localization of labeled Py with (C) Rab5-containing vesicles or (D) Rab7-containing vesicles at the early (0.5–2 hrs) and late (4–6 hrs) time points in both control and GD1a-supplemented NIH 3T3 cells. (E) Co-localization of labeled Py with Rab7-containing vesicles at 1–2 hrs post-infection in the ganglioside-deficient C6 cells. (F) Co-localization of labeled Py with LAMP1-containing vesicles 4–6 hrs post-infection in NIH 3T3 cells. (G) The extent of Py infection in GD1a-supplemented cells expressing wild type YFP-Rab7 or dominant negative YFP-Rab7 (DN). At least 220 transfected cells were analyzed from three independent experiments. All data are the mean+/−SD. A two-tailed t test was used.