Figure 4.
Figure 4: The H3K4me3/2 engagement by NUP98–JARID1A perturbs
the epigenetic state of developmentally critical loci during
haematopoiesis. a, The impact of mutations on Flag–NJL
binding to HOXA9 in 293 cells. WT, wild type. b, Immunoblot of
haematopoietic progenitors 10 days after transduction of vector,
wild-type or mutant NJL. Phosphorylated c-Kit (P-c-Kit) is a
marker of mast cells. Actin is shown as a loading control. c,
ChIP for Hoxa9 promoter-associated NUP98-fusion proteins (3 Flag-tagged)
and H3K4me3 in marrow progenitors 10 days after transduction. d,
e, Transforming capacities after introducing mutation to NJL (d)
or those by NUP98–PHF23 (e) or after replacing JARID1A-PHD3
with another PHD finger that engages either H3K4me3/2 or
H3K4me0. The total progenitor number was counted at day 1, 10,
25 and 40. f–h ChIP for Suz12 (f), Mll2-binding to Hoxa9/a11
(g), and Hoxa9-associated H3 acetylation (h) in marrow
progenitors 15 days after transduction of vector or NJL. Error
bar indicates s.d.; n = 3; *P < 0.05, **P < 0.005, ***P < 10^-4
and *****P < 10^-6. i, A scheme showing that NUP98–PHD fusion
acts as a boundary factor and prevents the spreading of polycomb
factors from Hoxa13/a11 to Hoxa9, thus inhibiting H3K4me3
removal and H3K27me3 addition during haematopoiesis.
The above figure is reprinted
from an Open Access publication published by Macmillan Publishers Ltd:
Nature
(2009,
459,
847-851)
copyright 2009.
Flag-tagged)
and H3K4me3 in marrow progenitors 10 days after transduction. d,
e, Transforming capacities after introducing mutation to NJL (d)
or those by NUP98–PHF23 (e) or after replacing JARID1A-PHD3
with another PHD finger that engages either H3K4me3/2 or
H3K4me0. The total progenitor number was counted at day 1, 10,
25 and 40. f–h ChIP for Suz12 (f), Mll2-binding to Hoxa9/a11
(g), and Hoxa9-associated H3 acetylation (h) in marrow
progenitors 15 days after transduction of vector or NJL. Error
bar indicates s.d.; n = 3; *P < 0.05, **P < 0.005, ***P < 10^-4
and *****P < 10^-6. i, A scheme showing that NUP98–PHD fusion
acts as a boundary factor and prevents the spreading of polycomb
factors from Hoxa13/a11 to Hoxa9, thus inhibiting H3K4me3
removal and H3K27me3 addition during haematopoiesis.