Fluorinated Fucose Derivative Impacts Rotavirus Infection

Briefly, 2F-peracetyl-fucose (2F) is a cell-permeable fluorinated fucose derivative that acts as an inhibitor of fucosyltransferases following its uptake and metabolic transformation into a GDP-fucose mimetic [44 (link),45 (link),46 (link)]. We used the protocols described previously [7 (link)]. For 2F (Sigma-Aldrich, US) treatment, 500 μM of 2F or dimethyl sulfoxide (DMSO, negative control) was added to differentiated PIEs and incubated for 3 days at 37 °C before they were infected with RVC as described above (fresh 2F/DMSO were added daily). Cell viability was examined using Trypan blue before infection to ensure no DMSO-associated toxic or other biological effects on PIEs. The infected PIEs were incubated for 2 days before harvesting. For sialidase treatment, MA104 cells in 96-well plates or PIEs were pre-treated with 10 mU (diluted in 50 mM Tris-HCl pH 7.5, 150 mM NaCl, 10 mM CaCl₂, and 0.02% NaN₃: TNC buffer) of sialidase/neuraminidase (NA) from Arthrobacter ureafaciens (Roche, US) for 1 h at 37 °C before inoculation. TNC buffer was used as a negative control for sialidase treatment. Following sialidase treatment, the PIEs were infected with RVCs and harvested as described above.