Figures & data
Figure 1 Overexpression of FUT7 increased the sLeX expression.
Notes: (A) Western blot analysis was used to assess the protein levels in A549 cells of forced expression of FUT7. (B) The expression of sLeX induced by overexpression of FUT7 was tested by immunocytochemistry.
Figure 2 Overexpression of FUT7 stimulated the colony formation and cell proliferation in A549 cells.
Notes: (A) The colony formation assay was conducted with A549-vector and A549-FUT7 cDNA cells. Colonies formed during 10 days of culture were visualized by crystal violet staining. The number of colonies formed in each group was calculated from three individual experiments. ***P<0.001. (B) Cell cycle analysis – A549-vector and A549-FUT7 cDNA cells were stained for DNA by PI; the stained cells were analyzed by FACScan flow cytometer using a ModFit 5.2 computer program. (C) Western blot to detect PCNA expression – total protein from the whole-cell lysate of A549-vector and A549-FUT7 cDNA cells was subjected to Western blot analysis using PCNA antibody.
Abbreviation: PI, propidium iodide.
Abbreviation: PI, propidium iodide.
Figure 3 Overexpression of FUT7 promoted invasion and migration in A549 cells.
Notes: (A) Cell migration was detected by wound-healing assay. (B) Cell invasion was examined by transwell (Matrigel-coated membrane). (C) Analysis of MMP2 and MMP9 activity by gelatin zymography. *P<0.05.
Figure 4 The impact of FUT7 overexpression and anti-sLeX antibody blocking on EGFR/Akt/mTOR signaling activation.
Notes: (A) P-EGFR, EGFR, p-Akt, Akt, p-mTOR and mTOR were detected by Western blot. (B) Immunoblot after immunoprecipitation of EGFR from cell lysates using an anti-sLeX antibody coupled to magnetic beads.
Figure 5 Involvement of Akt/mTOR signaling pathway in FUT7 overexpression promoted cell growth.
Notes: (A) Inhibitory effects of LY294002 on A549 cells were tested by CCK-8 assay. (B) The expression of p-Akt, Akt and PCNA after treatment of FUT7 cDNA and LY294002 alone or in combination. (C) The inhibitory effect of LY294002 on colony formation of A549 cells was tested by colony formation assay. The number of colonies formed in each group was calculated from three individual experiments. **P<0.01.
