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Fig. 6. Cell-autonomous activation of Krox20 by Hoxb1. (A-C)
Flat mount, at the level of the mid-hindbrain boundary, of a chick hindbrain
electroporated with a construct expressing HA-tagged Hoxb1, in situ
hybridized for Krox20 and immunolabelled for Hoxb1-HA: (A)
Krox20 staining, (B) anti-HA immunofluorescence and (C) merge.
(D-F) Transverse sections, at the level of the forebrain-midbrain, of
5-somite stage vhnf1-/- embryos co-injected with
meis1.1 and hoxb1aMyc RNAs, in situ hybridized for
krox20 and immunolabelled for Hoxb1aMyc: (D) krox20
staining, (E) anti-Myc immunofluorescence and (F) merge. (G) ChIP of
Hoxb1a-bound element C. Chromatin extracted from zebrafish embryos co-injected
with meis1.1 and Myc-tagged hoxb1a RNAs was subjected to the
ChIP procedure in the absence (first lane) or presence (second lane) of
anti-Myc antibody. Immunoprecipitated DNA was analyzed by PCR using primers
designed to amplify the core of element C (297 bp) or an unrelated sequence
located -22.4 kb upstream of the transcription start site (422 bp). The third
lane shows the PCR products obtained from the input chromatin.
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