0.01 0.10 1.00 10.00 100.000

20

40

60

80

100

120

0.01 0.10 1.00 10.00 100.000

20

40

60

80

100

120

cell

nu

mb

er (

% v

ehic

le c

on

tro

l)

cell

nu

mb

er (

% v

ehic

le c

on

tro

l)

cisplatin concentration (µM)gefitinib concentration (µM)

101 102 103

cell

nu

mb

er

expression level(fluorescence intensity)

control EGFR

sensitive

resistant

P-EGFR

total EGFR

P-AKT

total AKT

GAPDH

S2 R2S R

P-ERK 1/2

total ERK 1/2

cell

nu

mb

er

0.0 0.1 1.00

25

50

75

100

concentration of gefitinib (M)

% c

ells

0.0 0.1 1.00

25

50

75

100

concentration of gefitinib (M)

% c

ells

gefitinib concentration (µM) gefitinib concentration (µM)

% c

ells

% c

ells

0.0 0.1 1.0 0.0 0.1 1.0

100

75

50

25

0

100

75

50

25

0

sensitive resistant

Supplementary Fig. S1 Box et al.

A) B)

C)

E)

D)

F)

tum

ou

r vo

lum

e (c

m3)

Supplementary Fig. S2 Box et al.

0 40 80 120 16010

100

1000

10000

7 9 11 13 15 17 19 210.0

0.1

0.2

0.3

0.4

0.5A) B)

time (days)

cell

nu

mb

er (

% 0

h)

time (hours)

Supplementary Fig. S3 Box et al.

0

50

100

150

200

250

300

350

400

450

S R S2 R2g

efit

inib

-tre

ated

tu

mo

ur

volu

me

(% d

ay 0

)

A)

450

400

350

300

250

200

150

100

50

0

B)

0 3 7 10 140

50

100

150

200

250

300

350

400

time (days of therapy)CALS CALR CALS2 CALR2

gef

itin

ib-t

reat

ed t

um

ou

r vo

lum

e(%

veh

icle

-tre

ated

)

TSP-1

Serpin

B5

PD-ECGF

Angiogen

in

DPPIV

EG-VEGF

Coagula

tion fa

ctor I

II

TSP-2

CXCL16

Endothel

in-1

IGFBP-3

Angiost

atin

Angiopoie

tin-2

VEGF

Serpin

E1

uPAM

MP-9

HB-EGF

Amphire

gulin

EGFAct

ivin

A

Basic

FGF

Pentra

xin 3

IL-8

TIMP-1

-6

-4

-2

0

2

4

6

Supplementary Fig. S4 Box et al.

A)

B)

PF4IL

-1B

Prola

ctin

GM-C

SF

0.0

0.2

0.4

0.6

0.8

1.0

1.2

Sensitivity

Resistance

CALR only

β

exp

ress

ion

leve

l(a

rbit

rary

un

its)

Lo

g2 ra

tio

of

resi

stan

t/se

nsi

tive

exp

ress

ion

dmso gefi0

5000

10000

15000

20000

25000

30000

35000

Supplementary Fig. S5 Box et al.

A)

IL-8

GM-CSF

TIMP-1

dmso gefi0

10000

20000

30000

40000

50000

60000

dmso gefi0

1000

2000

3000

4000

5000

6000

7000

con

cen

trat

ion

(p

g/m

l)co

nce

ntr

atio

n (

pg

/ml)

con

cen

trat

ion

(p

g/m

l)

**

C)

E)

dmso gefi0

100200300400500600700800900

*

con

cen

trat

ion

(p

g/m

l)IL-8

dmso gefi0

5000

10000

15000

20000

25000

con

cen

trat

ion

(p

g/m

l)D)

B)

F)

dmso gefi0

20000

40000

60000

80000

100000

120000

TIMP-1

con

cen

trat

ion

(p

g/m

l)

GM-CSF

S2 R2 S2 R2

con gefitinib

S2 R2 S2 R2

con gefitinib

S2 R2 S2 R2

con gefitinib

PJS PJR PJS PJR

con gefitinib

PJS PJR PJS PJR

con gefitinib

PJS PJR PJS PJR

con gefitinib

Supplementary Fig. S6 Box et al.

# Maps pValue1 Inflammatory mechanisms of pancreatic cancerogenesis 1.945E-17 11 672 EGFR signaling pathway in Lung Cancer 6.244E-13 8 473 IGF signaling in HCC 1.480E-12 8 524 ERBB family and HGF signaling in gastric cancer 2.039E-12 8 545 Role of stellate cells in progression of pancreatic cancer 3.361E-10 7 606 Muscle contraction_Regulation of eNOS activity in endothelial cells 5.364E-10 7 647 Immune response_Oncostatin M signaling via MAPK in mouse cells 6.340E-10 6 358 Immune response_Oncostatin M signaling via MAPK in human cells 9.044E-10 6 379 Angiogenesis in HCC 6.023E-09 6 50

10 Development_Leptin signaling via JAK/STAT and MAPK cascades 8.628E-09 5 2511 Inhibition of apoptosis in pancreatic cancer 1.356E-08 6 5712 Development_Role of IL-8 in angiogenesis 1.510E-08 6 5813 Immune response_IL-17 signaling pathways 1.860E-08 6 6014 Development_PDGF signaling via STATs and NF-kB 3.224E-08 5 3215 Mucin expression in CF via IL-6, IL-17 signaling pathways 4.437E-08 5 3416 EGFR family signaling in pancreatic cancer 7.865E-08 6 7617 LKB1 signaling pathway in lung cancer cells 1.178E-07 5 4118 Immune response_IL-1 signaling pathway 1.697E-07 5 4419 Role of DNA methylation in progression of multiple myeloma 1.697E-07 5 4420 Immune response_Oncostatin M signaling via JAK-Stat in mouse cells 1.963E-07 4 1821 Role and regulation of Prostaglandin E2 in gastric cancer 2.382E-07 5 4722 IL-6 signaling in multiple myeloma 2.949E-07 5 4923 Cytokine production by Th17 cells in CF (Mouse model) 2.949E-07 5 4924 Immune response_Oncostatin M signaling via JAK-Stat in human cells 3.095E-07 4 2025 Pro-inflammatory action of Gastrin in gastric cancer 3.269E-07 5 5026 PI3K signaling in gastric cancer 3.617E-07 5 5127 Mucin expression in CF via TLRs, EGFR signaling pathways 3.617E-07 5 5128 Immune response_MIF-mediated glucocorticoid regulation 4.653E-07 4 2229 PGE2 pathways in cancer 5.314E-07 5 5530 Dual role of TGF-beta 1 in HCC 6.732E-07 4 2431 Effect of H. pylori infection on gastric epithelial cell proliferation 6.956E-07 5 5832 Development_Prolactin receptor signaling 6.956E-07 5 5833 Development_EGFR signaling pathway 1.056E-06 5 6334 Regulation of VEGF signaling in pancreatic cancer 1.105E-06 4 2735 Immune response_CD40 signaling 1.143E-06 5 6436 Immune response_IL-6 signaling pathway 1.489E-06 4 2937 DNA damage_Brca1 as a transcription regulator 1.715E-06 4 3038 Transcription_Ligand-dependent activation of the ESR1/SP pathway 1.715E-06 4 3039 EML4/ALK fusion protein in nonsmoking-related lung cancer 2.241E-06 4 3240 Influence of multiple myeloma cells on bone marrow stromal cells 2.545E-06 4 3341 IGF-1 signaling in pancreatic cancer 3.641E-06 4 3642 HBV signaling via protein kinases leading to HCC 3.641E-06 4 3643 Development_Beta-adrenergic receptors transactivation of EGFR 4.074E-06 4 3744 Cytokine production by Th17 cells in CF 5.053E-06 4 3945 Immune response_MIF in innate immunity response 5.603E-06 4 4046 Immune response_Neurotensin-induced activation of IL-8 in colonocytes 6.834E-06 4 4247 Development_ACM2 and ACM4 activation of ERK 7.519E-06 4 4348 Development_VEGF signaling and activation 7.519E-06 4 4349 K-RAS signaling in pancreatic cancer 8.254E-06 4 4450 Cell adhesion_Chemokines and adhesion 1.049E-05 5 100

Ratio

patient number5 6 7 9 10 11 13 14 15 16 17 18 19 20 22 23 24 25 27 28 30 31 32 34 35

0

50

100

150

200

250

Supplementary Fig. S7 Box et al.

IL-8

TIMP-1

con

cen

trat

ion

(p

g/m

l)co

nce

ntr

atio

n (

ng

/ml)

A)

B)

patient number5 6 7 9 10 11 13 14 15 16 17 18 19 20 22 23 24 25 27 28 30 31 32 34 35

0

100

200

300

400

500

600

700

800

900

1000

5 6 7 9 10 11 13 14 15 16 17 18 19 20 22 23 24 25 27 28 30 31 32 34 350

500

1000

1500

2000

2500

3000

Supplementary Fig. S7 (contd.) Box et al.

VEGF-AC)

con

cen

trat

ion

(p

g/m

l)

patient number

Supplementary Fig. S8 Box et al.

A)

B)

no rash rash0

20

40

60

80

100P

FS

(m

onth

s)

no rash rash0

20

40

60

80

100

OS

(m

onth

s)