Fig. 3

Fut8 participates in macrophage migration induced by ox-LDL. A The fucosylation level of the total protein of Raw 264.7 cells was determined by lectin blot after treatment with (0-75) mg/ml ox-LDL for 24 h. B Lectin blotting assays conducted to characterize the levels of total protein α-2,6 sialylation (SNA) and α-2,6 sialylation (MALI) levels in Raw 264.7 cells subjected to (0-75) mg/ml ox-LDL for 24 h. C The mRNA levels of Fut4, Fut7 and Fut8 in Raw 264.7 cells were analyzed by real-time RT-PCR after treatment with 0-75 mg/ml ox-LDL for 24 h. D The levels of total protein α-1,2-fucosylated (UEA1), α-1,3/4-fucosylated (LTL) and α-1,6-fucosylated (LCA) of Raw 264.7 cells were determined by lectin blot after treatment with 0-75 mg/ml ox-LDL for 24 h; E Western blotting was used to detect the expression of Fut8 protein in Raw 264.7 cells and mouse peritoneal macrophages after treatment with ox-LDL for 24 h. F,G Raw 264.7 cells were pretreated with siFut8 or pcDNA3.1-Fut8 plasmid, and then the number of cells on the dark side was identified by Transwell assay upon ox-LDL (50 μg/mL) treatment (bar=100 μm), the wound healing assay was used to identify the cell migration speed (bar=500 μm) or (H) the cells were stained for F-actin using Phalloidin-iFluor 488, and nuclei were counterstained with DAPI. Green:F-actin staining; Blue: DNA staining (bar=20 μm). Data were reported as the mean ± S.D. ( n=4, ^*p< 0.05 ; ^***p< 0.001, t-test.).