Axl glycosylation mediates tumor cell proliferation, invasion and lymphatic metastasis in murine hepatocellular carcinoma

Figure 1 Expression profile of Axl glycoprotein in mouse hepatocellular carcinoma cell lines. A: Axl glycoprotein levels by Western blotting analysis in Hca-P and Hca-F cell lines. Relative signal intensities of Axl protein were compared with GAPDH by LabWorks (TM ver4.6, UVP; BioImaging Systems), (^aP < 0.05 vs untreated Hca-F cells); B: Hca-F cells were treated with 0 μg/mL, 5 μg/mL, 10 μg/mL, and 20 μg/mL tunicamycin for 12 h. Total protein extracts were loaded for each sample; C: Hca-F cell protein was deglycosylated with 12 units of PNGase F in lysis buffer. Probes were incubated at 37 °C in a time-dependent manner (0 h, 8 h, 16 h, 24 h). Protein was separated on a gel for Western blotting Analysis. After sodium dodecyl sulfate-polyacrylamide gel electrophoresis, proteins were transferred to a polyvinylidene fluoride membrane, and were detected by rabbit anti-mouse Axl polyclonal antibody. GAPDH blotting was used as the control. GAPDH: Glyceraldehyde-3-phosphate dehydrogenase.