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Fig. 3. Barhl2 induces apoptosis in Xenopus neuroectoderm. (A) Stage 15
embryos. (a) Embryo injected with Xbarhl2AS and immunostained using
anti-phospho-histone H3 antibody. (b,c) In situ hybridization using
Xsox3 as a probe on embryos injected with: (b) Xbarhl2AS and
treated with HU; (c) human BCL2 (500 pg). (B) TUNEL staining of stage
15 embryos representative of those injected with mouse Barhl2 (20 pg)
(a), Xbarhl2 (20 pg) (b) and mouse Barhl2 (10 pg) (c). (a,b)
Anterior views, dorsal upwards; (c) dorsal view, anterior upwards. (C) Number
of TUNEL+ cells were counted on the injected side and compared to
the number on the control side at stage 15. The results were assessed by
Student's t-test. Results are shown as mean±s.e.m. Injection
of mouse Barhl2 increased apoptosis 2.6 times (10 pg to 25 pg)
(n=70, P=3.10–8), as does Xbarhl2
(20 pg, n=20, P=9.10–5). Injection of human
BCL2 (500 pg) reduced apoptosis (n=35, P=0.002),
and the effect of Barhl2 (10 pg) was reversed by co-injection of
human BCL2 (500 pg) (n=40, P>0.01). Injection of mouse
Barhl2
FIL had no effect (n=28, P>0.01).
(D) In situ hybridization using Xsox3 as a probe of stage 14 embryos
injected with: (a) mouse Barhl2 (10 pg), 82%; (b) human BCL2
(500 pg) 89%; and (c) mouse Barhl2 (10 pg) and human BCL2
(500 pg), 60%.
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