Science - USA (2022-05-27)

Tanget al., Science 376 , eabe1505 (2022) 27 May 2022 9of13

WANG_YAP_TAZ_TARGETSNES = -1.79

FDR = 0.001

Enrichment score (ES)

D

F

FOSL1 KD TAZ KD YAP KD YAP/TAZ KD

CRPC-SCL peaks CRPC-AR peaks

CRPC-AR peaks

CRPC-SCL peaks

A C

B YA P/TAZ KD in MSKPCa3

VINCULIN

YAP

TAZ

FOSL1

NT ctrlYAP

KD

TAZ

KD

YA P

/TAZ

KD

01234

0

2

4

6

MSKPCa3 (CRPC-SCL)

Days

RLU (normalized to day 0)

NT ctrl

YAP KD

TAZ KD

YAP/TAZKD

siDeath

********

**

Chromatin accessibility of MSKPCa3 upon knockdown

GSEA of MSKPCa3 upon knockdown

Coverage on

peaks (bpm)

YA PTAZCTGFCYR6^1

AJUBAANKRD1CCND

1

FOSL1

0.0

0.5

1.0

1.5

2.0

Relative expression

qPCR uponYA P/TAZ KD in MSKPCa3 (48h)

01234

0.0

0.5

1.0

1.5

2.0

2.5

Days

RL

U(

normalized to day 0)

-2.0 Peak center2.0Kb

0.0

0.2

0.4

Coverage on peaks (bpm)

-2.0 Peak center2.0Kb -2.0 Peak center2.0Kb -2.0 Peak center2.0Kb -2.0 Peak center2.0Kb

-2.0 Peak center2.0Kb -2.0 Peak center2.0Kb -2.0 Peak center2.0Kb -2.0 Peak center2.0Kb -2.0 Peak center2.0Kb

0.0

0.1

0.2

0.3

0.4

Control

FOSL

1 KDTAZ KDYAP KD

YAP/

TAZ KD

-30%

-20%

-10%

0%

10%

20%

% of chromatin accessibilitychange compared to contro

l

***

***

***

Chromatin accessibility change

compared to control

CRPC-AR peaks

CRPC-SCL peaks WANG_YAP_TAZ_TARGETSNES = -1.86

FDR = 0.003

Enrichment score (ES)

WANG_YAP_TAZ_TARGETSNES = -1.86

FDR = 0.002

Enrichment score (ES)

WANG_YAP_TAZ_TARGETSNES = -1.67

FDR = 0.04

Enrichment score (ES)

Enrichment score (ES)

FOSL1 KD TAZ KD YAP KD YAP/TAZ KD

E

** **

*

***

**** * **

*

** **** *

*

MSKPCa2 (CRPC-AR)

NT ctrl

YAP KD

TAZ KD

YAP/TAZKD

siDeath

NT ctrl

YAP KD

TAZ KD

YAP /TAZ KD

Fig. 6. Impact ofFOSL1, YAP, TAZ,andYAP/TAZknockdown.(A)qPCR
showing that double knockdown ofYAP/TAZby siRNA in MSKPCa3 leads to
decreased expression of downstream target genes andFOSL1. Mean ± SD,
n = 3. Nontargeting (NT) siRNA serves as the negative control. **P <
0.0001, P <0.01,*P < 0.05 (two-tailed unpairedt test); KD, knockdown.
(B) Western blot (WB) confirming the knockdown efficiency and the decrease
ofFOSL1expression72hoursaftertransfectioninMSKPCa3.(C)Cell
growth curves of MSKPCa3 and MSKPCa2 after siRNA knockdown. Mean ± SD,

n =8forMSKPCa3,n =4forMSKPCa2.****P <0.0001,**P <0.01(two-

tailed unpairedt test). (D) Chromatin accessibility changes uponFOSL1, TAZ,

YAP,andYAP/TAZknockdown at CRPC-ARand CRPC-SCL peaks. (E)Average

percentage of chromatin accessibility changes uponFOSL1, TAZ, YAP,and

YAP/TAZknockdown compared to control at CRPC-AR and CRPC-SCL peaks.

***P < 0.001 (permutation test). (F) GSEA plots showing negative enrichment

of YAP/TAZ target genes in MSKPCa3 upon knockdown ofFOSL1, TAZ,

YAP,andYAP/TAZ.

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