Derivation of fMaSC- and fSTR-Specific Gene
Expression Signatures
We performed microarray expression analyses on the fMaSC,
fSTR, and aMaSC populations to ascertain molecular pathways
with potential relevance to fetal mammary development, fMaSC
biology, and breast cancer. We obtained reproducible expres-
sion profiles from independent biological pools representing
each population and identified differentially expressed genes
comprising fMaSC, fSTR, and aMaSC signatures (Figure 3A
and Table S4; significance analysis of microarrays; false
discovery rate (FDR) < 10%; [Tusher et al., 2001]). We identified
869 unique genes more highly expressed in the fMaSC popula-
tion (the fMaSC signature) than in the fSTR and 812 unique genes
more highly expressed in the fSTR population (the fSTR signa-
ture) than in the fMaSC. Among the fMaSC signature genes,
34% were common to both the fMaSC and aMaSC popula-
tionswhen compared to fSTR, but40%were significantly over-
expressed in the fMaSC relative to the aMaSC (Figure 3A).
We confirmed the differential gene expression patterns
between the fMaSC and fSTR populations on a panel of genes
selected from putative stem cell, developmental, and cancer
relevant pathways (Figures 3B and 3C). Furthermore, high-
throughput single-cell qRT-PCR analyses confirmed expression
of a partially overlapping selection of 46 genes in individual cells
of the fMaSC population (Figure 3D). This approach also verified
that individual fetal cells coexpress luminal keratins, myoepithe-
lial keratins, and vimentin (Figure 3D).
Derivation of fMaSC- and fSTR-Specific Gene
Expression Signatures
We performed microarray expression analyses on the fMaSC,
fSTR, and aMaSC populations to ascertain molecular pathways
with potential relevance to fetal mammary development, fMaSC
biology, and breast cancer. We obtained reproducible expres-
sion profiles from independent biological pools representing
each population and identified differentially expressed genes
comprising fMaSC, fSTR, and aMaSC signatures (Figure 3A
and Table S4; significance analysis of microarrays; false
discovery rate (FDR) < 10%; [Tusher et al., 2001]). We identified
869 unique genes more highly expressed in the fMaSC popula-
tion (the fMaSC signature) than in the fSTR and 812 unique genes
more highly expressed in the fSTR population (the fSTR signa-
ture) than in the fMaSC. Among the fMaSC signature genes,
34% were common to both the fMaSC and aMaSC popula-
tionswhen compared to fSTR, but40%were significantly over-
expressed in the fMaSC relative to the aMaSC (Figure 3A).
We confirmed the differential gene expression patterns
between the fMaSC and fSTR populations on a panel of genes
selected from putative stem cell, developmental, and cancer
relevant pathways (Figures 3B and 3C). Furthermore, high-
throughput single-cell qRT-PCR analyses confirmed expression
of a partially overlapping selection of 46 genes in individual cells
of the fMaSC population (Figure 3D). This approach also verified
that individual fetal cells coexpress luminal keratins, myoepithe-
lial keratins, and vimentin (Figure 3D).
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