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Fig. 6. Endothelial cell proliferation is suppressed in
Man1GT/GT cells. (A,B)
Immunohistochemistry of EBs, grown in suspension for 10 days then explanted
for 6 days, using an antibody to PECAM-1. (A) Endothelial cells form extensive
branches in the wild-type (+/+). (B) The Man1GT/GT
explants form fewer and less organized branches. (C) FACS analysis of
PECAM-1+ cells isolated from Man1GT/GT and
wild-type EB explants, showing that in cultures of both genotypes the
percentage of PECAM-low expressing cells was similar, but the percentage of
PECAM-high expressing cells was strongly reduced in the
Man1GT/GT cultures. (D) TGFß1 further inhibits
cell proliferation in Man1GT/GT fibroblasts. Wild-type and
Man1GT/GT MEFs and SVEC cells were treated with TGFß1
for 72 hours at the indicated concentrations. Mean±s.e.m. (n=6
wells per treatment). Asterisks indicate statistically significant decrease in
the proliferation of Man1GT/GT MEFs compared with
wild-type and SVEC cells. (E) Real-time gene expression analysis of EB
explants. EBs were grown in suspension for 14 days then explanted for 4 days.
RNA for each data point was derived from >20 EBs and assayed in triplicate.
Data are shown as a ratio of Man1GT/GT relative to wild
type and were normalized to an average of S18 and Gapdh.
Mean±s.e.m.; *, P<0.05; **,
P<0.005; Student's t-test. Scale bar: 100 µm.
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