Figure 3.

Menaquinone-4 (MK4) altered gene expression of DT-diaphorase and osteoprotegrin detected at the proteome level. Rat aortic vascular smooth muscle cells (VSMCs) were cultured as described in MATERIALS AND METHODS for 24 hours in medium containing 1) 10% CDMEM + 25 µM MK4 (MK4) or 2) 10% CDMEM + no additions (control). After 24 hours the cells were continued growing for 6 hours in serum free medium containing 1) 25 µM MK4 and 2) no additions (control). DT-diaphorase activity was measured in the adherent cells as dicumarol sensitive DCPIP reducing activity (panel A). Osteoprotegrin secreted from control and MK4 treated cells are shown in panel B as Western blots which also represent total secreted osteoprotegrin (see MATERIALS AND METHODS). Standard deviations estimated for the measured DT-diaphorase activities are indicated on the top of the bars (n=3). The P-value was P<0.0001