Fig. 5.

Acquirement of tumour necrosis factor (TNF)-α suppressing activity in supernatants and intracellular adenosine triphosphate (ATP) depletion in time. (a) Whole blood was diluted 1:1 in normal phenol-red free (PRF) medium (filled bars) and in supernatants of untreated human umbilical vein endothelial cells (HUVEC) (Sup-untreated) or in supernatants of dopamine-treated HUVEC (Sup-Da-treated), obtained directly after 6 h (open bars) or 12 h (hatched bars) of cold storage. To each condition 0·5 μg/ml of lipopolysaccharide was added. TNF-α production was assessed as described in Fig. 3. A total of three different experiments were performed. The result of a representative experiment is expressed as mean TNF-α production (pg/ml) ± standard deviation (s.d.). (b) Untreated (open bars) and dopamine-treated (filled bars) HUVEC were stored at 4°C for different periods of time. Hereafter, intracellular ATP was measured as described in Materials and methods. The total amount of ATP before cold storage was taken as 100% for each condition. The result of a representative experiment is depicted and expressed as mean percentage of total ATP ± s.d. A total number of three experiments were performed.