The Influence of Ouabain on Human Dendritic Cells Maturation

Table 2

Na,K-ATPase activity.

	Activity (nmol/mg/min)
Ct	267.5 (±15.3)
Ouabain 100 nM (addition during; membrane preparation)	71.3 (±5.7)
Ouabain 100 nM (48 h)	111.0 (±3.1)

Results are the mean (±SD) of three independent determinations. ATP hydrolysis reaction was started by the addition of 3 mM γ³²P-ATP (sp. act 1520 cpm/nmol); after 40 minutes all reactions were stopped by the addition of 0.2 mL of 0.4 M perchloric acid + 0.4 mL of activated charcoal and then centrifuged. Supernatants from these centrifugations contained ³²Pi generated from ATP hydrolysis and specific counts were estimated through LSC in a Beckman 2100 counter. All measurements in each condition were discounted from the blanks (with acid denatured enzyme) and from the activity observed in the presence of 1 mM Ouabain to assess specific Na,K-ATPase activity. The results are the pool of cells from two donors.