Fig. 7. Differential expression of novel capillary morphogenesis genes (CMGs)
during morphogenesis in 3D collagen matrices. Differential display and cDNA
library screening were used to identify differentially expressed genes during
capillary morphogenesis. DNA sequencing revealed whether genes were known or
novel and gene-specific primers were prepared to assess differential
expression by RT-PCR (A). RT-PCR was performed to assess the mRNA expression
patterns of various CMGs and known genes over a 48 hour time course. Total RNA
was prepared from EC cultures at 0, 8, 24 and 48 hours of culture. The RNA was
reverse transcribed for RT-PCR and PCR was performed using gene-specific
primer sets. In addition, cDNA probes were made in select cases to perform
northern blots (B). The far-left panel in B represents a methylene
blue-stained blot indicating the position of the 28S and 18S rRNA bands and
showing equal loading of 3 µg of total RNA per lane. (C) A western blot of
cell extracts from a time course of capillary morphogenesis is shown using
affinity purified CMG-2 antibodies. MCH, melanin concentrating hormone;
GCPKRK, germinal center protein kinase related kinase-1.
