Figure
2.
HCR
inhibits
dendrite
formation
of
PIG1
and
MelanA
cells.
Representativephotoes
of
cell
morphology
of
PIG1
(A)
and
MelanA
(C)
cells.
PIG1
cells
were
pretreated
with
20
nM
ET-1
for
24h,
then
treated
with
different
concentrations
of
HCP
for
24h.
Cytoskeleton
was
visualized
by
cell
immunofluorescent
staining
for
F-actin.
Scale
bars =
20
µ
The
number
of
dendrites
were
counted
in
PIG1
(B)
and
MelanA
(D)
cells.
(E)
Melanin
secretion
to
the
medium
in
MelanA
cells
treated
with
20
nM
ET-1
in.the
presence
or
absence
of
HCP.
(F)
Quantitative
analysis
of
melanin
content
in
the
medium.
Data
were
expressed
mean
±
SD
from
3
independent
experiments
(n
=
3).
*P
0.05,
**P
0.01,
ET-1
vs
control
#P
0.05,
##P
0.01,
HCP
versus
ET-1.
164 JOURNAL OF COSMETIC SCIENCE

Figure
3.
HCR
inhibits
mlanin
transfer-related
gene
expression.
PIG1
(A)
and
MelanA
(D)
cells
were
pretreated
with
20
nM
ET-1
for
24h,
then
treated
with
different
concentrations
of
HCP
for
24h.
Rab27a
and
Rac1
levels
were
determined
by
Western
blot
analysis.
Relative
expression
levels
of
Rab27a
and
Rac1,
normalized
to
the
expression
of
GAPDH,
were
quantified
by
Image
J
software
in
PIG1
(B)
and
MelanA
(E)
cells.
The
expression
of
Rac1
mRNA
in
PIG1
(C)
and
MelanA
(E)
cells
treated
with
ET-1
or
HCP
were
quantified
by
using
real-time
PCR
analysis.
Data
were
expressed
mean
±
SD
from
3
independent
experiments
(n
=
3).
*P
0.05,
***P
0.001
ET-1
vs
control
#
p
0.05,
##
p
0.01,
###
p
0.01,
HCP
versus
ET-1.
165 Hydrolyzed Conchiolin Protein Inhibits Melanin