(a) p53 target gene expression in Chd7+/+ and
Chd7−/− (whi/whi) NCCs.
Averages+/−s.e.m. normalized to β-actin (n=5).
*,**p-values<0.05,<0.01, one-tailed Mann-Whitney test. (b) Top:
5′ end of p53 locus. Bottom: Representative of triplicate ChIP
analyses of Chd7 binding at p53 promoter (P1) and negative control open-chromatin regions
(P2 and P3), relative to input. +/−s.d. of technical triplicates. (c)
Bottom: p53 immunoblot of human fibroblasts derived from unaffected and CHARGE patients.
(Below: p53 levels after normalization to Actin.) Top: Averages+/−s.d of p53
levels. *p-value=0.015, one-tailed Student’s t-test. (d) p53 target
gene expression in low-serum treated human fibroblast cells derived from unaffected and
CHARGE patients relative to untreated samples. 5 CHARGE cell lines analyzed in duplicate.
Representative cell line pair is shown, +/−s.d. of technical triplicates.
(e) Left, p53 immunohistochemistry on thymi from fetuses with CHARGE
syndrome and unaffected fetuses. Arrows: p53-positive cells. Right, The number of
p53-positive cells per ten fields at 400× magnification, from one section.
(f) Comparison to control and
Chd7−/−p53+/+ embryos reveals
partial rescue of E10.5
Chd7−/−p53+/− embryos.
(Chd7−/− p53+/+: n=3;
Chd7−/−p53+/−: n=6).
(g) Embryo somite number for each genotype. ns=non-significant,
**,***p-value=0.0024,0.00032, two-tailed Student’s t-test.