Immunofluorescence analysis of A549 cells (Fig 5C), following infection with SPN, revealed that although R6:Ply-H exhibited a high degree of association with Gal8 (26.78 ± 3.17% at 10 h post infection (h.p.i)) (Fig 5D), R6:Ply-NH failed to co-localize with Gal8 at all the time points, confirming their residence inside intact vacuoles. This evidence concerns the gene SPN and infection.