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Fig. 1. Isolation of Pax2-expressing mid-hindbrain cells from mouse
embryos. (A) Structure of the Pax2GFP BAC transgene, which
was previously described as transgene 30 containing an in-frame GFP
insertion in exon 2 of Pax2
(Pfeffer et al., 2002). (B)
GFP expression of the Pax2GFP transgene in the
mid-hindbrain boundary (mhb) region, otic placode (op) and tail bud (tb) of
E8.5 embryos. (C) Representative FACS sorting of Pax2-expressing
mid-hindbrain cells. The mid-hindbrain domain (indicated by a bracket in B)
was manually dissected from a control wild-type and transgenic
Pax2GFP Pax2–/– embryo at E8.5
(seven somites) and dissociated into single cells by trypsin digestion. Live
PI– GFP+ cells were isolated from individual
embryos by fluorescence-activated cell sorting (FACS). The percentage of
GFP+ cells located within the sorting gate is shown for control and
transgenic embryos. PI, propidium iodide.
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