Transcript Powerpoint - Blood Journal

Disruption of Smad5 gene leads to enhanced proliferation
of high-proliferative potential precursors during embryonic
hematopoiesis
by Bing Liu, Yanxun Sun, Feizi Jiang, Shuangxi Zhang, Ying Wu, Yu Lan, Xiao Yang,
and Ning Mao
Blood
Volume 101(1):124-133
January 1, 2003
©2003 by American Society of Hematology
Increased frequency and replating potential of HPP-CFCs from Smad5−/− yolk sacs at E9.0 to
9.5.Dispersed yolk sac cells were cultured in HPP-CFC medium and the number of macroscopic
colonies was counted after 14 days of culture (A).
Bing Liu et al. Blood 2003;101:124-133
©2003 by American Society of Hematology
Increased incidence of hematopoietic EBs from Smad5−/− ES cells.ES cells were plated into
methylcellulose medium containing KL and IL-11 to form EBs. Note the extending hematopoietic
halo surrounding the central cell mass of 85% Smad5−/− EBs (A), whereas ap...
Bing Liu et al. Blood 2003;101:124-133
©2003 by American Society of Hematology
Increased BL-CFC number and altered expression of hemangioblast-related genes within day 3.5
Smad5−/− EBs.(A) Day 3.5 EBs were dissociated and then plated in BL-CFC assay culture
containing VEGF and KL. After 5 days of growth, 3 types of colonies, secondary...
Bing Liu et al. Blood 2003;101:124-133
©2003 by American Society of Hematology
Investigation of embryonic globin gene expression by RT-PCR within day 3.5 and 4.0 EBs.The
arrows on the top of each photo represent the 10-fold dilution of samples.
Bing Liu et al. Blood 2003;101:124-133
©2003 by American Society of Hematology
Identification of Ery/Ps within wild-type and Smad5−/− EBs.(A) EBs were dissociated at day 4 to
6 and replated into cultures containing Epo favoring the growth of primitive erythroid colonies.
Bing Liu et al. Blood 2003;101:124-133
©2003 by American Society of Hematology
Identifications of HPP-CFC and primitive progenitor-related gene expression within day 6 wildtype and Smad5−/− EBs.HPP-CFCs derived from day 6 EBs of wild-type (A) and Smad5−/− (B) ES
cells were able to generate macroscopic colonies in vitro.
Bing Liu et al. Blood 2003;101:124-133
©2003 by American Society of Hematology
Reduced sensitivity of Smad5−/− HPP-CFCs to TGF-β1 inhibition.The inhibitory effect of TGF-β1,
added to HPP-CFC assay at final concentrations of 0.03, 0.1, 0.3, and 2 ng/mL, is shown.
Bing Liu et al. Blood 2003;101:124-133
©2003 by American Society of Hematology
Secondary and tertiary colonies derived from primary wild-type and Smad5−/− HPP-CFCs in
replating assay.Most of the Smad5−/− HPP-CFC colony was able to form secondary HPP-mix
consisting of erythrocytes, granulocytes, mast cells, and macrophages (B,F,J), whe...
Bing Liu et al. Blood 2003;101:124-133
©2003 by American Society of Hematology
Altered gene expression within Smad5−/−HPP-CFCs.Macroscopic HPP-CFC colonies derived
from day 6 wild-type and Smad5−/− EBs were individually plucked and mixed for RT-PCR
examination.
Bing Liu et al. Blood 2003;101:124-133
©2003 by American Society of Hematology