TGFβig-h3 IGF-I - + - IGF-II - - + Supplementary Figure 1. Stimulated release of TGFβig-h3 by IGF-I and IGF-II.

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TGFβig-h3 IGF-I - + - IGF-II - - + Supplementary Figure 1. Stimulated release of TGFβig-h3 by IGF-I and IGF-II. Western blots shows both IGF-I (50ng/ml) and IGF-II (100ng/ml) stimulated TGFβig-h3 release from gastric myofibroblasts. Cells GAPDH Cells TGFβig-h3 Media TGFβig-h3 Ionomycin - + Supplementary Figure 2. Calcium-dependent stimulated release of TGFβig-h3 from gastric myofibroblasts. A) Western blot showing that IGF-II (100 ng/ml) stimulated release of TGFβig-h3 was decreased in Ca2+ free media. B) The calcium ionophore, ionomycin (1µM), stimulated TGFβig-h3 release from gastric myofibroblasts. 37kDa 68kDa 74kDa A B - + - + - - + + IGF-II Ca2+ free

MMP-7 MMP-7 + AG1024 F/Fo 1.20 0.95 Supplementary Figure 3. Intracellular Ca2+ transients induced by MMP-7 are inhibited by AG1024. MMP-7 (2 g/ml) increased intracellular calcium in myofibroblasts (top) and this was inhibited in the presence of AG1024 (3.2µM) (bottom); records from a region of interest placed in the centre of the cell.

A. Decorin B. Galectin-3 Sequence: SAIQLGNY Sequence: IALDFQR Sequence IGF-II Mean ratio IGF-II vs control Quantified in (out of 3 replicates) DLPPDTTLLDLQNNK 1.26 3 SAIQLGNYK 0.99 VPGGLAEHK 1.51 2 VVQCSDLGLDK 1.18 VVQCSDLGLDKVPK 0.43 Sequence: SAIQLGNY B. Galectin-3 Sequence IGF-II Mean ratio IGF-II vs control Quantified in (out of 3 replicates) IALDFQR 0.77 3 Sequence: IALDFQR

Mean ratio IGF-vs control Quantified in (out of 3 replicates) C. TGFig-h3 Sequence IGF-II Mean ratio IGF-vs control Quantified in (out of 3 replicates) EGVYTVFAPTNEAFR 2.50 3 GDELADSALEIFK 1.73 LTLLAPLNSVFK 1.58 2 STVISYECCPGYEK 3.04 VLTDELK 2.65 Sequence: EGVYVTVFAPTNEAFR

Mean ratio IGF-II vs control Quantified in (out of 3 replicates) D. IGFBP-5 Sequence IGF-II Mean ratio IGF-II vs control Quantified in (out of 3 replicates) AVYLPNCDR 1.55 3 FVGGAENTAHPR 2.10 GVCLNEK 1.29 HMEASLQELK 1.35 IISAPEMR 1.58 QESEQGPCR 1.51 2 Sequence: AVYLPNCDR Supplementary Figure 4. Representative examples of SILAC data for identification and quantification of peptides from media of responder cells. The examples shown are A) decorin, b) galectin-3, C) TGFig-h3 and D) IGFBP-5. In each case, tryptic peptides shown underlined on a yellow background were quantified in at least 2 out of 3 replicate samples; representative spectra for one peptide are shown, together with a Table of the mean ratio of abundance in IGF-II treated samples compared with control.

A. Preprosecretogranin II 10 20 30 40 50 60 MAEAKTHWLG AALSLIPLIF LISGAEAASF QRNQLLQKEP DLRLENVQKF PSPEMIRALE 70 80 90 100 110 120 YIENLRQQAH KEESSPDYNP YQGVSVPLQQ KENGDESHLP ERDSLSEEDW MRIILEALRQ 130 140 150 160 170 180 AENEPQSAPK ENKPYALNSE KNFPMDMSDD YETQQWPERK LKHMQFPPMY EENSRDNPFK 190 200 210 220 230 240 RTNEIVEEQY TPQSLATLES VFQELGKLTG PNNQKRERMD EEQKLYTDDE DDIYKANNIA 250 260 270 280 290 300 YEDVVGGEDW NPVEEKIESQ TQEEVRDSKE NIEKNEQIND EMKRSGQLGI QEEDLRKESK 310 320 330 340 350 360 DQLSDDVSKV IAYLKRLVNA AGSGRLQNGQ NGERATRLFE KPLDSQSIYQ LIEISRNLQI 370 380 390 400 410 420 PPEDLIEMLK TGEKPNGSVE PERELDLPVD LDDISEADLD HPDLFQNRML SKSGYPKTPG 430 440 450 460 470 480 RAGTEALPDG LSVEDILNLL GMESAANQKT SYFPNPYNQE KVLPRLPYGA GRSRSNQLPK 490 500 510 520 530 540 AAWIPHVENR QMAYENLNDK DQELGEYLAR MLVKYPEIIN SNQVKRVPGQ GSSEDDLQEE 550 560 570 580 590 600 EQIEQAIKEH LNQGSSQETD KLAPVSKRFP VGPPKNDDTP NRQYWDEDLL MKVLEYLNQE 610 KAEKGREHIA KRAMENM Patient 1, shotgun (8 tryptic peptides). Patient 2, shotgun (8 tryptic peptides). B. Preproenkephalin 10 20 30 40 50 60 MARFLTLCTW LLLLGPGLLA TVRAECSQDC ATCSYRLVRP ADINFLACVM ECEGKLPSLK 70 80 90 100 110 120 IWETCKELLQ LSKPELPQDG TSTLRENSKP EESHLLAKRY GGFMKRYGGF MKKMDELYPM 130 140 150 160 170 180 EPEEEANGSE ILAKRYGGFM KKDAEEDDSL ANSSDLLKEL LETGDNRERS HHQDGSDNEE 190 200 210 220 230 240 EVSKRYGGFM RGLKRSPQLE DEAKELQKRY GGFMRRVGRP EWWMDYQKRY GGFLKRFAEA 250 260 LPSDEEGESY SKEVPEMEKR YGGFMRF Patient 1, shotgun (1 hemi-tryptic peptide) Patient 2, shotgun (2 tryptic peptides, I hemi-tryptic) Patient 3, SILAC ± IGF (2 tryptic peptides)

C. Peptidyl α-amidating mono-oxygenase 10 20 30 40 50 60 MAGRVPSLLV LLVFPSSCLA FRSPLSVFKR FKETTRPFSN ECLGTTRPVV PIDSSDFALD 70 80 90 100 110 120 IRMPGVTPKQ SDTYFCMSMR IPVDEEAFVI DFKPRASMDT VHHMLLFGCN MPSSTGSYWF 130 140 150 160 170 180 CDEGTCTDKA NILYAWARNA PPTRLPKGVG FRVGGETGSK YFVLQVHYGD ISAFRDNNKD 190 200 210 220 230 240 CSGVSLHLTR LPQPLIAGMY LMMSVDTVIP AGEKVVNSDI SCHYKNYPMH VFAYRVHTHH 250 260 270 280 290 300 LGKVVSGYRV RNGQWTLIGR QSPQLPQAFY PVGHPVDVSF GDLLAARCVF TGEGRTEATH 310 320 330 340 350 360 IGGTSSDEMC NLYIMYYMEA KHAVSFMTCT QNVAPDMFRT IPPEANIPIP VKSDMVMMHE 370 380 390 400 410 420 HHKETEYKDK IPLLQQPKRE EEEVLDQGDF YSLLSKLLGE REDVVHVHKY NPTEKAESES 430 440 450 460 470 480 DLVAEIANVV QKKDLGRSDA REGAEHERGN AILVRDRIHK FHRLVSTLRP PESRVFSLQQ 490 500 510 520 530 540 PPPGEGTWEP EHTGDFHMEE ALDWPGVYLL PGQVSGVALD PKNNLVIFHR GDHVWDGNSF 550 560 570 580 590 600 DSKFVYQQIG LGPIEEDTIL VIDPNNAAVL QSSGKNLFYL PHGLSIDKDG NYWVTDVALH 610 620 630 640 650 660 QVFKLDPNNK EGPVLILGRS MQPGSDQNHF CQPTDVAVDP GTGAIYVSDG YCNSRIVQFS 670 680 690 700 710 720 PSGKFITQWG EESSGSSPLP GQFTVPHSLA LVPLLGQLCV ADRENGRIQC FKTDTKEFVR 730 740 750 760 770 780 EIKHSSFGRN VFAISYIPGL LFAVNGKPHF GDQEPVQGFV MNFSNGEIID IFKPVRKHFD 790 800 810 820 830 840 MPHDIVASED GTVYIGDAHT NTVWKFTLTE KLEHRSVKKA GIEVQEIKEA EAVVETKMEN 850 860 870 880 890 900 KPTSSELQKM QEKQKLIKEP GSGVPVVLIT TLLVIPVVVL LAIAIFIRWK KSRAFGDSEH 910 920 930 940 950 960 KLETSSGRVL GRFRGKGSGG LNLGNFFASR KGYSRKGFDR LSTEGSDQEK EDDGSESEEE 970 YSAPLPALAP SSS Patient 3, SILAC ± IGF (5 tryptic peptides)

D. Preproadrenomedullin 10 20 30 40 50 60 MKLVSVALMY LGSLAFLGAD TARLDVASEF RKKWNKWALS RGKRELRMSS SYPTGLADVK 70 80 90 100 110 120 AGPAQTLIRP QDMKGASRSP EDSSPDAARI RVKRYRQSMN NFQGLRSFGC RFGTCTVQKL 130 140 150 160 170 180 AHQIYQFTDK DKDNVAPRSK ISPQGYGRRR RRSLPEAGPG RTLVSSKPQA HGAPAPPSGS APHFL Patient 1, shotgun (1 hemi-tryptic peptide) Patient 3, SILAC ± IGF (2 tryptic peptides) 10 20 30 40 50 60 MLSCRLQCAL AALSIVLALG CVTGAPSDPR LRQFLQKSLA AAAGKQELAK YFLAELLSEP 70 80 90 100 110 NQTENDALEP EDLSQAAEQD EMRLELQRSA NSNPAMAPRE RKAGCKNFFW KTFTSC E. Preprosomatostatin 271, shotgun (1 tryptic, 1 hemi-tryptic peptide) Supplementary Figure 5. Examples of neuroendocrine markers identified in the secretomes of responder myofibroblasts. The sequences of A) preprosecretogranin II, B) preproenkephalin, C) peptidyl α-amidating mono-oxygenase (PAM), D) preproadrenomedullin (ADM), and E) preprosomatostatin are shown; in each case, quantified peptides are indicated by horizontal arrows. The data are from three different responder secretomes (patients 1 and 2 taken from Holmberg et al., J.Proteome Res., 2103; patient 3, this study). Validation of the expression of SGII and proenkephalin is provided in Fig 5. Validation of PAM, adrenomedullin and somatostatin localisation is shown by immunohistochemistry using Texas Red labelled secondary antibody, with SGII identified by FITC labelled secondary (PAM, ADM). Signal peptides in italics. Scale bars 10 m.

A B SGII SGII GAPDH GAPDH SGII-siRNA - + SGII-EP - + Supplementary Figure 6. Knockdown and overexpression of SGII in gastric myofibroblats. A) Representative Western blots showing decreased SGII abundance after siRNA knockdown. B) Representative Western blots showing increased SGII abundance after transfection with SGII expression plasmid (SGII- EP).