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There the cells were injected with a gene that makes green fluorescent protein, a standard way of marking cells.
By marking cells early in mouse development and then waiting for cancer to develop, the researchers simply looked inside the cancer to find the label and identify the original cell type.
"I think cancer biologists will start thinking about this because it is a more elegant way of marking cells than we currently use," says Leonard Zon, who directs the stem cell program at Boston's Children Hospital.
Live embryos were stained for apoptotic cells with the vital dye acridine orange (Sigma) that permeates acidic lysosomal vesicles and becomes fluorescent, thus marking cells dying by apoptosis.
This is consistent with our results, and supports the conclusion that Ascl1CreERT2 is marking cells that endogenously express Ascl1, even at low levels.
To quantify numbers of cells with epithelial and mesenchymal phenotypes in cultured cells, we employed flow cytometry and started by monitoring cells for the epithelial marker EpCAM as well as monitoring for vimentin or CD44 marking cells with mesenchymal attributes.
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Cells were rinsed with PBS + 3%FBS and DAPI was added for marking cell viability.
We identified distinct signatures that marked cells destined for positive selection versus apoptotic deletion.
First, diverse DNA barcodes can be used to uniquely mark cells and can be easily read out by sequencing.
Many of the genes targeted ES cells in KOMP are designed to mark cells with reporter molecules and produce conditional alleles.
Thus, the co-expressing GFP marks cells that express the UAS-x constructs.
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