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Fig. 4. Compensatory role of NKX6.2 for NKX6.1 function in 
- and ß-cell
development. Confocal images showing immunofluorescence detection of glucagon
(Glc) and insulin (Ins) in pancreas from wild-type, Nkx6.2-/-,
Nkx6.1-/- and
Nkx6.1-/-;Nkx6.2-/- embryos at E18.5 (A-D) and
E14.5 (G-J). Nkx6.2-/- (B,H) and wild-type (A,G) embryos
show similar numbers and organization of insulin+ and
glucagon+ cells in the pancreas. In Nkx6.1-/-
(I) and Nkx6.1-/-;Nkx6.2-/- (J) embryos,
glucagon+ cells are normal, but insulin+ cells are
reduced in number at E14.5. At E18.5,
Nkx6.1-/-;Nkx6.2-/- embryos (D) show a further
reduction in insulin+ cells as well as a reduction in
glucagon+ cells compared with Nkx6.1-/- embryos
(C). (E) Quantification of hormone+ cells in wild type and
Nkx6 compound mutants at E18.5. The average number of
glucagon+ and insulin+ cells per section was determined
by counting immunofluorescence-labeled cells. Five independent pancreata were
evaluated for each genotype and the average number of cells in wild type
arbitrarily set to 100. (F,K) Quantitative RT-PCR analysis of insulin and
glucagon in pancreata from Nkx6 compound mutants at E18.5 (F) and
E15.5 (K). Hormone expression levels are shown relative to the levels for
-actin mRNA from two independent measurements of four pancreata for
each genotype. For comparison, values are shown as a % of wild type, which was
arbitrarily set to 100%. At E18.5, insulin mRNA levels were 6.2±0.78%
of wild type in Nkx6.1-/- and 1.7±0.30% of wild
type in Nkx6.1-/-;Nkx6.2-/- embryos (F),
compared with 2.2±0.16% of wild type in Nkx6.1-/-
and 2.0±0.84% of wild type in
Nkx6.1-/-;Nkx6.2-/- embryos at E15.5 (K). The
values shown represent mean±s.e.m. *P<0.01, as
determined by Student's t-test. Scale bar: 50 µm.
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