Supplementary Figure 1

b

a

USP2a

-actin

No

tran

sf.

Lipo

Vec

tor

pUS

P2a

WT

48 h 72 h LNCaP

No

tran

sf.

Lipo

Vec

tor

pUS

P2a

WT

Vector (1) pUSP2aWT (2)

% A

popt

osis

(su

b-G

1)

Contr CDDP DXT Doxo

1 2 1 2 1 2 1 2Contr CDDP DXT Doxo

cPARP

-actin

LNCaP

3 4 3 4 3 4 3 4Contr CDDP DXT Doxo

siContr (3)siUSP2a (4)

Contr CDDP DXT Doxo

LNCaP

cPARP

-actin%

Apo

ptos

is (

sub-

G1)

d

c

USP2a

-actin

48 h 72 h

No

tran

sf.

Lipo

siC

ontr

siU

SP

2a

No

tran

sf.

Lipo

siC

ontr

siU

SP

2a

LNCaP

Supplementary Figure 1 USP2a triggers chemo-resistance in LNCaP prostate cancer cells.

(a, c) Evaluation of USP2a protein expression, and (b, d) apoptosis analysis by sub-G1 and PARP cleavage (cPARP) carried out in drug-treated LNCaP cells undergoing transient transfection, as follows: without DNA or silencing oligos (Lipo), with pCDNA3 (Vector, 1), pCDNA3_USP2aWT (2), with siControl oligos, (3) or specific siUSP2a oligos (4). (e) Evaluation of drug response in the indicated LNCaP clones carried out by clonogenic assay.39

e

Sur

viva

l (%

)

CDDP (µg/ml) DTX (nM) Doxo (µM)

USP2aMUT

USP2aWT

EV

LNCaP

USP2aMUT

USP2aWT

EV

LNCaP

USP2aMUT

USP2aWT

EV

LNCaP