Discussion
We demonstrated that CMPs could effectively differentiate
into adipocytes via two TFs,
Klf4
and
c-Myc
, without
adipogenic stimulation. Neither
Klf4
nor
c-Myc
transduction
alone resulted in CMPs differentiating into adipocytes. These
results suggested that KM proteins in CMPs coordinately
regulate adipogenesis. Interestingly, this new protocol was
only effective in CMPs, but not in MSCs from bone marrow and
3T3-L1 preadipocytes. CMPs did not differentiate into adipo-
cytes when treated with adipogenic stimulation cocktails,
contradicting the hypothesis that CMPs contain adipogenic
Figure 6
In vitro
and
in vivo
IRI model. (
a
) Schematic representation of the
in vitro
IRI method. PBS, phosphate-buffered saline. Growth medium indicates the basal medium
for CMP. Hypoxia indicates 1% O
2
, 5% CO
2
, balance N
2
conditions in a hypoxic chamber, and normoxia indicates 21% O
2
, 5% CO
2
conditions. (
b
) Phase contrast microscope
images of CMPs under hypoxic conditions. (
c
) qRT-PCR analysis of the expression of each gene in CMPs. Error bars indicate S.E. (
n
=
3). * and ** indicate significant changes
compared with CMPs at 0 h (white box,
P
o
0.05 and 0.01, respectively). H indicates hypoxia condition; N indicates hormoxia condition. (
d
) Schematic representation of the
in vivo
IRI method. (
e
) Photograph of open-chest mouse with 8-0 Prolene suture thread on the LAD. (
f
) qRT-PCR analysis of the expression of each gene in an IRI model LV heart. Error
bars indicate S.E. (
n
=
3). * and ** indicate significance (
P
o
0.05 and 0.01, respectively).
‘
I
’
indicates ischemic condition;
‘
R
’
indicates reperfusion condition. (
g
) Western blotting
of IRI model LV heart. Error bars indicate S.E. (
n
=
3). * and ** indicates statistically significant differences at 0 h (white box,
P
o
0.05 and 0.01, in the order described)
Cardiac adiposity is regulated by
Klf4
and
c-Myc
D Kami
et al
8
Cell Death and Disease
