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Fig. 3. (A) Vegfa expression in early kidney development compared to
Wt1. (a,b) E12.5 kidneys, fixed immediately after dissection and
sectioned. (c,d) E11.5 metanephric kidney rudiments placed in culture for 48
hours. (a,c) Vegfa antisense probe, (b) Wt1 antisense probe
and (d) Vegfa sense probe. Clear Vegfa expression is present
in the condensed mesenchyme of the E12.5 kidney (a) overlapping significantly
with Wt1 expression at the same timepoint (b). (B) Vegfa stimulation
of branching and tubulogenesis. (a) Control PBS addition, (b) Vegfa addition,
Cytokeratin stain red, proximal tubule-binding lectin green. (c) Terminal
branch tip (P<0.001) and (d) proximal tubule (P<0.001)
quantitation showing results of four pairs of organ cultures represented by
(a) and (b), which are from the same embryo. (C) Antibody inhibition of
branching. (a) Control organ culture treated with rabbit IgG. (b) Organ
culture from the same embryo as (a) treated with anti-Vegfa neutralizing
antibody. (c,d) Quantitation of four pairs of organ cultures treated as in (a)
and (b); (c) terminal branch tips (P<0.001); (d) tubules as
identified by lectin staining for proximal tubules (P<0.001).
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