Epigenetic silencing of microRNA-203 is required for EMT and cancer stem cell properties Joseph H. Taube 1,2 , Gabriel G. Malouf 3,4 , Emily Lu 5 , Nathalie Sphyris 1,2 , Vidya Vijay 1,2 , Priyanka P. Ramachandran 1,2 , Katumasa R. Ueno 1 , Sanchaika Gaur 1 , Milena S. Nicoloso 6 , Simona Rossi 6 , Jason I. Herschkowitz 7 , Jeffrey M. Rosen 7 , Jean-Pierre J. Issa 8 , George A. Calin 6 , Jeffrey T. Chang 5,9 , Sendurai A. Mani 1,2, *
7
Embed
Epigenetic silencing of microRNA-203 is required for … A Figure S2 (Taube et al.) m i R - 2 0 3 l e t - 7 a m i R - 9 2 a m i R - 1 0 7 m i R - 1 2 6 m i R - 1 5 5 m i R-2 0 0 b
This document is posted to help you gain knowledge. Please leave a comment to let me know what you think about it! Share it to your friends and learn new things together.
Transcript
Epigenetic silencing of microRNA-203 is required for EMT and cancer stem cell
properties
Joseph H. Taube1,2, Gabriel G. Malouf3,4, Emily Lu5, Nathalie Sphyris1,2, Vidya Vijay1,2,
Priyanka P. Ramachandran1,2, Katumasa R. Ueno1, Sanchaika Gaur1, Milena S.
Nicoloso6, Simona Rossi6, Jason I. Herschkowitz7, Jeffrey M. Rosen7, Jean-Pierre J.
Issa8, George A. Calin6, Jeffrey T. Chang5,9, Sendurai A. Mani1,2, *
Figure S1 (Taube et al.)
a
b
miR-92b
HMLETwist
0.0
0.5
1.0
1.5
2.0
2.5
Rel
ativ
e Ex
pres
sion
miR-375
HMLETwist
0.0
0.5
1.0
1.5
Rel
ativ
e Ex
pres
sion
miR-1247
HMLETwist
0.0
0.5
1.0
1.5
2.0
2.5
Rel
ativ
e Ex
pres
sion
miR-1281
HMLETwist
0.0
0.5
1.0
1.5
Rel
ativ
e Ex
pres
sion
miR-203
miR-203
HMLETwist
0.0
0.5
1.0
1.5
Rel
ativ
e Ex
pres
sion
Figure S1 – Expression of other microRNAs with changes in promoter DNA methylation
Expression of indicated microRNAs was determined in HMLE control and HMLE Twist cells by
Taqman quantitative PCR. Levels were normalized to sno-U6.
Rel
ativ
e Ex
pres
sion
Vehicle 5azaC0.0
0.5
1.0
1.5
2.0
Rel
ativ
e Ex
pres
sion
Vehicle 5azaC0
5
10
15
Rel
ativ
e Ex
pres
sion
Vehicle 5azaC0
2
4
6
8
10
Rel
ativ
e Ex
pres
sion
Rel
ativ
e Ex
pres
sion
Vehicle 5azaC0.0
0.5
1.0
1.5
2.0
Rel
ativ
e Ex
pres
sion
Vehicle 5azaC0.0
0.5
1.0
1.5
2.0
miR-210
miR-92b miR-375 miR-1247miR-1281 miR-210
Rel
ativ
e Ex
pres
sion
HMLETwist
0
10
20
30
ns ns ns ns
nsnsnsnsns
Vehicle 5-AzaC0
5
10
15
p < 0.0001
p = 0.0212 p = 0.0058
D
A
Figure S2 (Taube et al.)
miR-20
3let
-7a
miR-92
a
miR-10
7
miR-12
6
miR-15
5
miR-20
0b
miR-22
4
miR-45
0a
miR-87
40.00001
0.0001
0.001
0.01
0.1
1
10
100 SUM159 ControlSUM159 miR-203
Fold
Cha
nge
Figure S2 – Expression of other microRNAs is unchanged by miR-203 over-expression
Relative levels of indicated microRNAs were quantified by Taqman quantitative PCR from RNA
extracts of control and miR-203 expressing SUM159 cells. Levels were normalized to sno-U6
and graphed relative to let-7a in control cells.
Figure S3 (Taube et al.)
a b
Figure S3 – miR-203 expression does not significantly affect cell growth rates
Cells were counted after the specified number of days in culture for both HMLE Twist (a) and
SUM159 (b) cells with either a control vector or miR-203.
Hs578t cells with either a control vector or expressing miR-203 were plated for mammosphere formation and spheres which formed after 10 days were counted.