Supplementary Figure 1
Supplementary Figure 2
Supplementary Figure 3
A
B
A B
C
Supplementary Figure 4
Supplementary Figure 5
A
B
AREG
HBEGF
EGF
ErbB3
EREG
TGFα
BTC
ErbB2
Mouse secondary
Goat secondary
Supplementary Figure 6
7
Days
Co
nce
ntr
atio
n u
g/m
L
0.1
1
10
100
0 5 10 15 20 25 30
B07
0.1 mg/kg1 mg/kg10 mg/kg
Supplementary Figure 7
10mg/kg Nonspecific IgG 10mg/kg MEDI3622
tEGFR
tErB3
ErB2(Tyr1112)
ErB2(Thr686)
EGFR(Tyr1086)
EGFR(Tyr845)
EGFR(Ser1070)
EGFR(Tyr1173)
Supplementary Figure 8
Nonspecific IgG #1
MEDI3622 #2MEDI3622 #1
Nonspecific IgG #2
Yes
Src
Supplementary Figure 9
Supplementary Figure 10
Supplementary Figure 11
Supplementary Figure 12
A
B
Supplementary Figure 12
C
Supplementary Figure 13A B
C D
Supplementary Figure 14
Supplementary Table 1Cell line
% Max Inhibition MEDI3622
% Max Inhibition cetuximab
786-0 0.2 0.3
A549 3.4 6.1
BxPC3 16 3
C2BBE1 19.7 12
CakiI 19.2 25.6
Cal27 20.7 21.7
CCK81 8.7 77.5
Detroit 32.6 32.9
Du145 13.8 9.9
Fadu 16.5 35.2
H292 33.4 53.8
H358 41.5 65.9
HSC3 0 4.8
HT29 0 5.1
KYSE30 5.2 7.7
KYSE450 7.6 43.3
NUGC4 65.8 91.5
OE21 25 33.9
OE33 0.5 0.6
panc0203 16.4 5
PLCPRF5 30.9 48.2
SCC4 32.1 40.7
SCC9 34.5 58.6
SW48 31.8 49.6
SW403 36.2 43
T84 13.6 14.6
colo205 2 4
dld1 1 3
H650 3 3
HCT8 5.5 1
kato3 2.5 3.5
LS1034 0 5.4
PC3 2.2 0
Correlation of MEDI3622 and cetuximab anti-proliferative activity. Indicated cell lines were incubated with 400 nM MEDI3622 or 200 nM cetuximab to determine maximum inhibition of each antibody as indicated in Materials and Methods. Percent inhibition was determined on day 4 using Cell Titer Glow. Results for each cell line are indicated.
Supplementary Table 2fold decrease in protein shedding gene
7.817 NQO13.3399 PNPO2.6787 DPCD2.3452 RPUSD22.307 TPP12.0503 PTPRF2.0349 COG41.8526 PDXDC11.8002 SPINT11.7911 DLD1.7397 EXOSC101.7384 ACBD61.7291 PTK71.711 DAK1.6936 GPX21.6754 RBM31.6535 TXNRD11.6204 SEMA4D
Identification of ADAM17 substrates in COLO205 cells by SILAC proteomics. COLO205 cells at 75% confluence were preincubated with either 400 nM MEDI3622 or 400 nM nonspecific IgG. ADAM17 was then activated by addition of 50 nM PMA for 6 hours and cell supernatants were combined and analyzed by mass spectroscopy as described in Materials and Methods. Highlighted genes are shared with OE21 SILAC results.
fold decrease in protein
shedding gene4.955401 PTK74.881859 DESP3.691127 AREG3.483835 GSN3.255738 MRC23.106941 MAP2K13.105494 GPC12.933756 PTPRG2.858531 PODXL22.807885 UBLA42.806151 PVRL12.778241 ZNF2922.683267 SEMA4C2.54868 PTPLAD1
2.539231 SEMA4D2.388973 MERTK2.314226 SORL12.222173 PRRC12.148228 MUC162.128973 PTPRS2.088991 XRN2
2.043945 PTPRF
fold decrease in protein
shedding gene2.024742 GPC42.005254 PWP21.986176 NRP11.972114 SDSL1.831032 SDC21.791858 CP1.787981 PHGDH1.738012 F11R1.716561 amigo21.713238 notch21.678106 HLA-A1.676502 ST141.671877 JaG11.668446 NES1.663395 MUC11.636233 DAG11.621587 GEG11.618359 LXN1.618044 VLDLR1.595405 LRP11.58985 ALCAM
1.568062 ULBP2
Supplementary Table 3
Identification of ADAM17 substrates in H358 lung cancer cells by SILAC proteomics. H358 cells at 75% confluence were preincubated with either 400 nM MEDI3622 or 400 nM nonspecific IgG. ADAM17 was then activated by addition of 50 nM PMA for 6 hours and cell supernatants were combined and analyzed by mass spectroscopy as described in Materials and Methods.