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At 0.5 and 2 h post uptake, ∼34% and ∼54% of the virus was judged to be co-localised with LAMP-1.
Nevertheless, figure 7 shows that at 0.5 h and 2 h post uptake, BTV-1 was co-localised with co-internalised Alexa-568 labelled dextran, which also suggest that BTV-1 may use macropinocytosis or a macropinocytosis-like mechanism to enter BHK cells.
It's believed that α-synuclein null mice are protected against MPTP intoxication because lack of α-synuclein expression affects a yet to be defined process that occurs post uptake of MPP+ into the terminal and pre-mitochondrial complex I inhibition [42].
The majority of sifA mutants lose their vacuolar membrane by 8 h post uptake (Beuzón et al., 2000) and therefore provide a positive control for vacuolar membrane loss.
At 2 h and 16 h post uptake in each cell type, the growth of the steC mutant was indistinguishable from that of the wild-type strain, while the ssaV mutant displayed a strong replication defect in both cell types (data not shown).
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This difference was visible at 1 day post-uptake (Fig. 1B).
Similarly, at 15 minutes post-uptake, virus was not co-localised with LAMP-1 (data not shown).
This ca.25% increase in post-uptake metabolism between peptides and their amino acid monomers strongly suggests that there was no extracellular cleavage of peptides prior to uptake.
Invasin is a bacterial surface protein that interacts with mammalian β1-integrins and causes uptake of the bacterial cells, while listeriolysin O is a hemolysin that disrupts the endosomal membrane and allows bacteria to enter the mammalian cytoplasm post-uptake.
To confirm the presence of intracellular S. aureus within hMDMs at various times post-uptake, cells were fixed and stained with acridine orange (AO) and examined by confocal microscopy.
However, the pattern of virus uptake by concanamycin-A treated cells (detected at 0.5 h post-uptake by confocal microscopy using the PM10 antibody) was indistinguishable from mock-treated cells (data not shown), indicating that the inhibitory effect of concanamycin-A on infection was not due to a failure of the virus to be internalised.
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com