*4.2. Sulfated Fucan*

Extraction: Freshly frozen LH was thawed, blended into small particles (≤ 3 mm), and kept in 90 ◦C H2O for 6 h. The solution was filtered (20–25 μm particle retention) using a vacuum pump; CaCl2 was added to the filtrate and it was filtrated again (5 μm particle retention). The newly obtained filtrate was purified through ultrafiltration (300 kDa MWCO) and lyophilized.

Hydrolyzation: Lower-molecular-weight samples were obtained by mild acid hydrolysis of the ultrapurified sulfated fucan at pH = 3.0, 70 ◦C, and 5 min for Fuc2 and 35 min for Fuc3. The hydrolysates were cooled in ice water, dialyzed again with distilled water for 3 h (12 kDa MWCO), and lyophilized.

The purified and dried LH sulfated fucans Fuc1, Fuc2, and Fuc3 were dissolved in 5 mg/mL Ampuwa bidest (Fresenius, Schweinfurt, Germany). Before testing, the extracts were filtered with 0.2 μm Sarstedt filters (Nümbrecht, Germany) and further diluted in adequate medium to the concentrations mentioned in 4.6, 4.7, 4.8 and 4.9.
