Figure 4 Western blotting analysis of de novo VEGF‐C protein synthesis. Human RPE cells were treated with VEGF 100 ng/ml, glucose 15 mM, VEGF‐A 100 ng/ml plus glucose 15 mM, or 15 mM mannitol for 24 hours. Conditioned media were collected and immunoprecipitated using a polyclonal antibody raised against VEGF‐C. The immunoprecipitates were electrophoresed on a 12% SDS‐PAGE followed by blotting on a nitrocellulose membrane. Positive bands were visualised by an ECL detection system. The band intensity was quantified by laser densitometry and the amount of VEGF‐C was normalised against the amount of α tubulin present. (A) Cells were exposed to VEGF 100 ng/ml, glucose 15 mM, or VEGF‐A 100 ng/ml plus glucose 15 mM. (B) Cells were exposed to glucose 15 mM or mannitol 15 mM. The vertical bars represent SEM; p value (*p<00.5, **p<0.01) indicates the significant different between the different treatments and untreated control.