Figure 5.
Effects of PLG on uPA-MKs. (A) Confocal images of day-11 CD34+ MKs preincubated for 24 hours with Alexa-488 scuPA (green), Alexa-568 FV (red), and Alexa-647 ncPLG (white). Nuclei were stained blue by DAPI. For quantitation of overlap, see also Table 1. Profile measurements are as in Figure 2A. (B) WB analysis of lysates prepared from day-11 CD34+ MKs with or without loading with enzymatically active PLG (20 μg/mL) on day 10 for 18 hours followed by a wash and incubation with scuPA or uPAT (200 nM each) and/or FV (400 nM) for 24 hours. WB membranes were probed with mouse monoclonal antibodies recognizing reduced FV, nonreduced uPA, and rabbit polyclonal antibody recognizing reduced VWF and β-actin (nonconjugated and HRP-conjugated, respectively).