*4.7. VEGF ELISA*

Secreted VEGF in ARPE-19 supernatant was determined with the Human VEGF DuoSet ELISA Kit from R&D Systems (Wiesbaden, Germany) according to manufacturer's protocol. Supernatants were collected after three days of stimulation with different fucoidan extracts and an MTT assay was

conducted to set the VEGF e ffect in relation to the cell viability as previously described [19]. The medium was exchanged 24 h before collecting of supernatant.
