suppl. figure 1
DESCRIPTION
Suppl. Figure 1. Pituitary Tumors. Normal. miR-107 levels. miR-145 levels. miR-103 levels. miR-143 levels. Normal. Pituitary Tumors. miR-125b levels. miR-15b levels. miR-141 levels. miR-16 levels. miR-144 levels. miR-186 levels. miR-164 levels. let-7b levels. - PowerPoint PPT PresentationTRANSCRIPT
Suppl. Figure 1
miR
-107
le
vels
Normal Pituitary Tumors
miR
-103
le
vels
miR
-125
b
leve
lsm
iR-1
41
leve
lsm
iR-1
44
leve
lsm
iR-1
64
leve
ls
miR
-145
le
vels
miR
-143
le
vels
miR
-15b
le
vels
miR
-16
leve
lsm
iR-1
86
leve
lsle
t-7b
le
vels
Pituitary TumorsNormal
Suppl. Figure 1. Evaluation of the expression levels of differentially expressed microRNAs between pituitary normal and cancer tissues. MicroRNA expression levels were assessed by real-time PCR analysis in 5 normal and 12 cancer pituitary tissues. The experiment has been performed in triplicate and data are shown as mean ± SD.
Suppl. Figure 2
UGUUAGGGAAUUUUACUUGAA
UGGAUAGGACUUAAUGAACUU
8-mer
3’UTR PTEN
miR-26b
PTEN1 32951278-84
3’ UTR
UGUGCAGCCACGUCACUGUGA
UUUCUCUGGCCAAGUGACACU
8-mer
3’UTR BMI1
miR-128
BMI11 1654459-65
3’ UTR
Suppl. Figure 2. Sequence complementarity between miR-26b and the 3’UTR of PTEN gene and between miR-128 and the 3’UTR of BMI1 gene.
Suppl. Figure 3
a
- + - - -as-miR NC
# o
f co
lon
ies
- - + + +as-miR-26b- - - + -siRNA NC- - - - +siRNA-PTEN
# o
f co
lon
ies
- + - - -as-miR NC- - + + +as-miR-128- - - + -siRNA NC- - - - +siRNA-BMI1
b
- + - - -as-miR NC- - + + +as-miR-128- - - + -siRNA NC- - - - +siRNA-BMI1
# o
f in
vad
ing
cel
ls
- + - - -as-miR NC- - + + +as-miR-26b- - - + -siRNA NC- - - - +siRNA-PTEN
# o
f in
vad
ing
cel
ls
Suppl. Figure 3. MiR-26b and miR-128 control the tumorigenicity and invasiveness of AtT-20 pituitary tumors cell through regulation of PTEN and BMI1, respectively. (a) Number of colonies (mean ± SD) and (b) invading AtT-20 cells untreated or treated with 50nM antisense-microRNA negative control (as-miR-NC), antisense-microRNA-26b (as-miR-26b), antisense-microRNA-128 (as-miR-128), siRNA negative control (siRNA NC) and siRNA against PTEN (siRNA-PTEN).
# o
f co
lon
ies
- + - - - -miR-NC- - + - - -as-miR-NC- - - + - +as-miR-26b- - - - + +miR-128
p<0.0001
# o
f in
vad
ing
cel
ls
- + - - - -miR-NC- - + - - -as-miR-NC- - - + - +- - - - + +miR-128
p<0.0001
as-miR-26b
Suppl. Figure 4a
d
PT
EN
mR
NA
leve
ls
- + - - - -miR-NC- - + - - -as-miR-NC- - - + - +as-miR-128- - - - + +miR-26b
e
b c
- + - - - -miR-NC- - + - - -as-miR-NC- - - + - +as-miR-128- - - - + +miR-26b
AK
T p
ho
sph
leve
ls
f
# o
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ies
- + + +- + + +- - + +- - - +
as-miR-26bmiR-128
siRNA NCsiRNA-PTEN
# o
f in
vad
ing
cel
ls
- + + +- + + +- - + +- - - +
as-miR-26bmiR-128
siRNA NCsiRNA-PTEN
Suppl. Figure 4. MiR-26 and miR-128 regulate the PTEN-AKT pathway in AtT-20 pituitary cells. (a) Number of colonies and (b) invading AtT-20 cells, untreated or treated with 50nM miR-NC, as-miR-NC, as-miR-26b and miR-128. (c) Fold enrichment of BMI1 in the promoter area of PTEN in AtT-20 cells treated with 50NM as-miR-NC or as-miR-128, assessed by chromatin immunoprecipitation followed by real-time PCR analysis. (d) PTEN mRNA expression levels (mean ± SD) assessed by real-time PCR analysis and (e) AKT phosphorylation levels (S473) in AtT-20 cells treated for 48h with 50nM as-miR-NC, as-miR-128, miR-26b and their combinations. (f) Number of colonies and invading AtT-20 cells, untreated or treated with 50nM as-miR-26b and miR-128 or combination of as-miR-26b, miR-128 and siRNA NC or combination of as-miR-26b, miR-128 and siRNA-PTEN. The experiments have been performed in triplicate and data are shown as mean ± SD.
Suppl. Figure 5
GH3
MtT/S
# o
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vad
ing
cel
ls
- + + +- + + +- - + +- - - +
as-miR-26bmiR-128
siRNA NCsiRNA-PTEN
Suppl. Figure 5. Number of invading GH3 and MtT/S cells, untreated or treated with 50nM as-miR-26b and miR-128 or combination of as-miR-26b, miR-128 and siRNA NC or combination of as-miR-26b, miR-128 and siRNA-PTEN. The experiments have been performed in triplicate and data are shown as mean ± SD.