Suppl. figure 1

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HEKU2O

ST47

DHela

MCF-7

MDA MB 23

1

MDA MB 43

5s

MDA MB 46

8

Rel

ativ

e no

rmal

ized

miR

-21

amou

nts

(HE

K=1

)

Suppl. figure 2

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pmiR

-21-luc

pmiR

-21-luc

+ LNA m

iR-21

pmiR

-21-luc

+ LNA sc

ramble

Luci

fera

se/R

enill

a ra

tiore

lativ

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con

trol

PTEN

Vinculin

100 101 101 86 102

Moc

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LNA

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mbl

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miR

-21

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urso

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lin-4

pre

curs

or

Suppl. figure 3

Suppl. figure 4

Vinculin

p53

100 109 96 83 56

Moc

k

LNA

miR

-21

LNA

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Suppl. figure 5

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Control siRNA PDCD4 siRNA

O.D

. val

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LNA scrambledLNA miR-21

Suppl. Fig. 1. Expression of miR-21 in cell lines. The amount of miR-21 and U6 RNA was quantified using Taqman MicroRNA Assays (Applied Biosystems). Shown are the relative normalized miR-21 amounts (HEK=1). Suppl. Fig. 2. Efficient inhibition of miR-21 by the LNA inhibitor. Firefly luciferase reporter assay with pmiR-21-luc containing perfect compementarity to the mature miR-21 sequence. pmiR-21-luc was cotransfected into MCF-7 cells along with a Renilla luciferase transfection control plasmid, either alone or together with a miR-21 inhibitor or a scrambled control. Shown are relative luciferase values normalized to transfections with the reporter alone. Suppl. Fig. 3. Western blot for PTEN using lysates from MCF-7 cells transfected with miR-21 or lin-4 precursors or inhibitory miR-21 or scrambled LNAs. The bands were quantified relative to the appropriate Vinculin loading controls using a LAS-3000 imager (Fuji) and the relative quantifications are shown. Suppl. Fig. 4. Western blot for p53 using lysates from MCF-7 cells transfected with miR-21 or lin-4 precursors or inhibitory miR-21 or scrambled LNAs. The bands were quantified relative to the appropriate Vinculin loading controls using a LAS-3000 imager (Fuji) and the relative quantifications are shown. Suppl. Fig. 5. Knockdown of PDCD4 does not significantly affect cellular proliferation. O.D. values from a MTT growth assay in MCF-7 cells treated with PDCD4 or control siRNAs and transfected with inhibitors against miR-21 or a scrambled control. Data are shown as the mean ± SD of 3 replicates and are representative of 3 independent experiments.