Fig. 5. Expression of the genes encoding the CD36/LIMPII receptor, galectin as well as ADAMTS in primmorphs in response to different cultivation matrices. (A) The dissociated cells (time 0) were transferred into a seawater-based medium and cultured in uncoated dishes for 6 days (free). Then the primmorphs were transferred onto galectin-coated dishes for either 3 days (total period of incubation: 9 days) or 6 (12) days during which time they attached (attached: arrow). In a parallel series of experiments the primmorphs remained in uncoated dishes where they remained in the free state for 6 (12) days (free). Furthermore, the primmorphs that had been cultured on galectin-coated dishes were co-incubated for 6 (12) days with 10 µg/ml of the recombinant SDADAMTS peptide, containing the CSVTCG motif [+TSP]. RNA was isolated from the respective primmorphs and the same amount of RNA was loaded onto the gels and size separated. After blot transfer the filters were probed with the CD36/LIMPII receptor cDNA. (B) In parallel, the blot was probed with the galectin cDNA. (C) Effect of MTN on primmorphs that had been transferred to galectin-coated dishes for 3 (9) days. Either no compound was added to the cultures, or MTN was added at a concentration of 0.03 or 0.01 µg/ml to the assays for this incubation period. The blot was developed with the CD36/LIMPII receptor cDNA. (D) In parallel, the blot was incubated with the SDADAMTS probe. (E) As a control to show the interaction between the recombinant SDADAMTS peptide and the CD36/LIMPII receptor, this peptide was pre-incubated with equal concentrations of the recombinant CD36/LIMPII receptor polypeptide as described under Materials and Methods (+TSP/rCD36). The primmorphs were incubated on galectin-coated dishes for 6 (total incubation period of 12) days either in the absence of any recombinant polypeptide, or in the presence of 10 µg/ml recombinant SDADAMTS (+TSP), or in the presence of 10 µg/ml recombinant SDADAMTS together with 10 µg/ml recombinant CD36/LIMPII receptor (+TSP/rCD36). The relative degree of expression was correlated with that seen at the related control time point (day 0 or day 6, as indicated); this value is set to 1.