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Fig. 7. Distribution of BrdU in rabbit ocular surface epithelial cells sorted by flow cytometry. (A-D) Representative images of the experiments with limbal and conjunctival cells sorted following a 10-day label infusion. Frames A and B are limbal SP and LSSC cells. Note that in both populations the percentile of BrdU+ cells (yellow-green fluorescence) is low. Images were captured using dual red-green emission filtering. Unlabeled cells are shown as inserts for reference purposes. Frames C and D belong to the adherent cell fractions (Methods) from conjunctival G0-G1 and SP cells, respectively. For this experiment, green and red emissions were captured independently and superimposed, as graphically depicted on the left-hand side of C. Note that in the G0-G1 fraction the majority of cells incorporate BrdU whereas only a few SP cells are BrdU-positive. (E-H) Images from an experiment where the rabbit was infused for the full pump duration (14-16 days) followed by a 38-day label chase. Frames F and E correspond to conjunctival G0-G1 and SP respectively. Frames G and H are from the non-LSSC and LSSC limbal cells. Small inserts are images of BrdU-free nuclei.
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