Interplay between de novo and salvage pathways of GDP-fucose synthesis
Fig 5
Overexpression of FCSK and FPGT in TSTA3KO and GMDSKO HEK293T cell lines.
(A) intracellular concentration of GDP-fucose of wild-type, TSTA3KO and GMDSKO HEK293T cells, and in cells with overexpressed FPGT and FCSK proteins (tagged with HA and cmyc, respectively) in knockouts. (B) quantification of the percentage of fucosylated structures in wild-type, TSTA3KO and GMDSKO HEK293T cells and in cells with overexpressed FPGT or FCSK proteins in knockouts. (C) western blotting of HA in GMDSKO and TSTA3KO cells and overexpressed FPGT and FCSK in cell lines deficient in TSTA3 and GMDS enzymes. (D) western blotting of fucokinase in GMDSKO and TSTA3KO cells and overexpressed FPGT and FCSK in cell lines deficient in TSTA3 and GMDS enzymes. As a loading control, the anti-HSP60 antibody was used. For (A) and (B), ns, not significant; *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001 as determined using one-way ANOVA with the Tukey post-hoc test. Data are represented as mean ± SEM. Each sample was run in three biological replicates.