L o g [R X C 0 0 5 ] (M )

% i

nh

ibit

ion

-1 4 -1 2 -1 0 -8 -6 -4

-2 0

0

2 0

4 0

6 0

8 0

1 0 0

1 2 0

B T K W T

B T K C 4 8 1 S

-1 1 -1 0 -9 -8 -7 -6 -5 -4

0

2 0

4 0

6 0

8 0

1 0 0

1 2 0

L o g [R X C 0 0 5 ] (M )

B T K W T

B T K C 4 8 1 S

% i

nh

ibit

ion

-1 2 -1 0 -8 -6 -4

0

2 0

4 0

6 0

8 0

1 0 0

1 2 0

lo g [R X C 0 0 5 ] (M )

% o

f a

cti

va

ted

B-c

ell

s

(Ig

M-s

tim

ula

ted

CD

19

+ C

D6

9+

ce

lls

)

-1 2 -1 0 -8 -6 -4

-2 0

0

2 0

4 0

6 0

8 0

1 0 0

1 2 0

lo g [R X C 0 0 5 ] (M )

% o

f a

cti

va

ted

B-c

ell

s

(Ig

D-s

tim

ula

ted

CD

19

+ C

D6

9+

ce

lls

)

pBTK

BTK

B-Actin

γ γ

α

Cell line

OCI-Ly10 (pIC50) 8.2± 9.3±0.2 8.3±0.1 8.4±0.2

TMD8 (pIC50) 8.3± 9.1±0.2 8.3 (N=2) 8.2 (N=2)

IgM stimulation

pBtk

gapdh

Btk

00251868 CLL

Veh

icle

20

0 n

MR

XC

00

5

50

0 n

MIb

ruti

nib

Veh

icle

DC

9 (

+ co

ntr

ol)

50

0 n

MR

XC

00

5

10

0 n

MR

XC

00

5

0 1 0 2 0 3 0 4 01 .0

1 .5

2 .0

D a y s p o s t la s t t r e a t m e n t

RT

V (

Re

lati

ve

Tu

mo

ur

Vo

lum

e)

****

***

***

* * * * p < 0 .0 0 0 1

* * * p < 0 .0 0 1

0 2 0 4 0 6 0

D a y s

Tu

mo

ur

vo

lum

e (

mm

3)

V e h ic le B ID

R X C 0 0 5 [1 0 m g / k g ] B ID

R X C 0 0 5 [5 0 m g / k g ] B ID

Ib r u t in ib [1 0 m g / k g ] Q D

2 0 0

3 0 0

4 0 0

5 0 0

1 5 0

1 0 0T re a tm e n t R e g r o w th

0 1 0 2 0 3 0

T r e a t m e n t t im e (D a y s )

Tu

mo

ur

vo

lum

e (

mm

3)

V e h ic le B ID

R X C 0 0 5 1 0 m g / k g B ID

R X C 0 0 5 5 0 m g / k g B ID

Ib r u t in ib 1 0 m g / k g Q D

2 0 0

3 0 0

4 0 0

5 0 0

1 5 0

1 0 0

****

* * * * p < 0 .0 0 0 1

***

* * * p < 0 .0 0 1

0 7 1 4 2 1

T r e a t m e n t t im e (d a y s )

Tu

mo

ur

vo

lum

e (

mm

3)

V e h ic le

R X C 0 0 5 3 0 m g / k g B ID

1 0 0

2 0 0

4 0 0

R X C 0 0 5 1 0 m g / k g B ID

8 0 0

1 0 0 0

* * * * p < 0 .0 0 0 1

****

****