|
|
|
|
|
|
|
||||
| Home Help Feedback Subscriptions Archive Search Table of Contents | ||||
|
Fig. 3. Temporal changes in Rnf2 association, H3-K9 acetylation and H3-K27
trimethylation at the Hoxb8 putative promotor region. (A)
The expression of Hoxb8 in the cranial and caudal tissues at various
developmental stages, as quantified by using real-time PCR against the
expression of Gapdh. Relative quantity was compared between anterior
and posterior tissues. The Hoxb8/Gapdh ratio in the anterior
tissue of 8.0 dpc embryo was arbitrarily 1. Embryos were bisected into the
anterior and posterior tissues as described below. (B) Six- and
eight-somite embryos were bisected at the level of the newly generated somite
boundary and separated into anterior segmented (A) and posterior unsegmented
(P) regions, and WCE prepared from respective tissues were subjected to ChIP
analyses. (C) Embryos at the 
13-somite stage were dissected into
three pieces at the boundary between somites 9 and 10, and at the newly
generated somite boundary. WCE prepared from cranial region up to somite 9 (A)
and posterior unsegmented region (P) were subjected to ChIP analyses.
(D) Embryos at 9.5 dpc (about 25 somites) were bisected into anterior
(A) and posterior (P) pieces at the level of somite 9/10 boundary and
respective WCE were subjected to ChIP analyses. (E) Embryos at 10.5 dpc
(35 somites) were dissected at the level of the posterior end of the
hindbrain and caudally to the forelimb bud after removing the viscera. For
ChIP using anti-Rnf2, chromatin fractions prepared from the anterior (A) and
posterior (P) tissues were used, whereas WCE were used for anti-acetylated
H3-K9 and -trimethylated H3-K27 antibodies. (F) Schematic comparisons
of Rnf2 association, H3-K9 acetylation and H3-K27 trimethylation at the region
3 between anterior and posterior tissues. The relative quantity of each
genomic region in immunoprecipitated genomic DNA from anterior and posterior
tissues was estimated by referring to those of `Input' DNA isolated from the
initial lysates and enrichment values against the `Input' are represented by
the black and gray bars, respectively. Stages left unexamined are indicated by
boxes crossed with a diagonal line.
|
