chip
assays
performed
terms
a20
iκbα
gapdh
promoters
using
antibodies
corresponding
primers
described
substantiate
finding
took
advantage
chromatin
immunoprecipitation
chip
assay
examine
whether
binding
nf-κb
endogenous
promoter
influenced
vivo
expression
uxt
diminished
consistently
deficiency
endogenous
uxt
level
resulted
considerable
decreases
amount
p65
associated
endogenous
cognate
promoters
upon
tnf-α
stimulation
given
uxt
interacted
p65
essential
maintain
presence
nf-κb
inside
nucleus
wondered
whether
uxt
integral
component
nf-κb
transcriptional
enhanceosome
vivo
address
possibility
transfected
ha-uxt
293t
cells
performed
systematic
chip
assays
promoters
a20
iκbα
described
materials
methods
turned
out
uxt
indeed
present
within
nf-κb
transcriptional
enhanceosome
notably
presence
became
much
more
prominent
upon
stimulation
suggested
uxt
dynamically
recruited
onto
enhanceosome
addition
uxt
nothing
transcription
complex
gapdh
promoter
indicating
selectivity
uxt
action
fig.
also
stimulated
293t
cells
performed
similar
chip
assays
confirm
again
endogenous
uxt
recruited
onto
nf-κb
enhanceosome
response
stimulation
fig.
ng/ml
tnf-α
stimulation
chip
assays
performed
a20
iκbα
gapdh
promoters
describedchromatin
immunoprecipitation
indicated
ddx3x
recruited
ifn
promoter
upon
infection
listeria
monocytogenes
suggesting
transcriptional
mechanism
action
ddx3x
found
tbk1
substrate
vitro
vivo
phosphorylation-deficient
mutants
ddx3x
failed
synergize
tbk1
ability
stimulate
ifn
promoter
overall
data
imply
ddx3x
critical
effector
tbk1
necessary
type
ifn
induction
address
question
used
chromatin
immunoprecipitation
chip
amplified
enhanceosome-binding
region
ifn-β
promoter
quantitative
pcr
figure
5a
irf3
absent
under
non-stimulated
conditions
recruited
ifn
promoter
upon
infection
l.
monocytogenes
figure
5b
specificity
signal
ascertained
using
control
serum
chip
chip
suggests
ddx3x
recruited
ifn
promoter
positioning
ddx3x
downstream
tbk1
level
irf3cells
cultured
24h
prepared
using
chip
assay
kit
from
upstate
biotechnology
inc.
lake
placid
ny
according
manufacturer
recommendations
preformed
previously
describe
primers
foxo3a
sco2
promoter
shown
supplemental
methods
location
shown
supplemental
fig
s3
ip
transient
transfection
ip
westerns
done
previously
described
bands
ips
detected
using
ecl
protocol
santa
cruz
biotechnology
santa
cruz
ca
visualized
fuji
las-3000
intelligent
darkbox
fujifilm
medical
systems
stamford
ct
chip
analysis
showed
wt-sirt3
cells
increase
foxo3a
binding
upstream
regulatory
regions
gene
promoters
each
contain
two
canonical
foxo3a
binding
sites
roughly
kb
upstream
transcription
start
site
supplemental
fig
s3
foxo3a
binding
both
mnsod
fig.
3d
upper
panel
sco2
lower
panel
promoters
increased
wt-sirt3
compared
mt-sirt3
cells
experiments
imply
sirt3
may
increase
foxo3a
dna-binding
chip
analysis
foxo3a
finding
mnsod
sco2
promoters
hct116
cells
overexpress
either
wild-type
deacetylation
null
sirt3
gene