Fig. 9. Alteration in SA lineage-specific transcription factor expression in purified adrenal chromaffin cells. (A) Single-cell suspensions dissociated from E18.5 adrenal glands were analyzed by flow cytometry after staining with FDG (a fluorogenic substrate for ß-galactosidase) and PE-conjugated CD45 antibody. lacZ⁺CD45^- (red quadrants) and lacZ^-CD45^- (blue quadrants) fractions were isolated. The percentages of cells in lacZ⁺CD45^- fractions are indicated. (B) The mRNA level of Sf1, a cortex-specific nuclear receptor, was undetectable in the lacZ⁺CD45^- fractions, indicating the high purity of chromaffin cells. (C) Gata3, Th, Dbh, Hand2, Phox2b and Mash1 mRNA levels in the lacZ⁺CD45^- fractions (normalized to GAPDH mRNA) were assayed by real-time RT-PCR. Data were collected from E18.5 individual embryos of Gata3^Z/- (5), Gata3^Z/-:Tg^hDBH-G3 (5) or Gata3^Z/+ (5) genotypes. Data are presented as mean±s.e.m. The statistical significance of the differences between Gata3^Z/- and Gata3^Z/+ are indicated (^*P<0.05; ^**P<0.01; Student's t-test).