Human Non-Small Cell Lung Cancer (H3255) Cell Line; Trypsin Digest; Antibody: Py; CST #9411 Treatments: Untreated (Heavy), Iressa-Treated (Light)

Human Non-Small Cell Lung Cancer (H3255) Cell Line; Trypsin Digest; Antibody: Py; CST #9411 Treatments: Untreated (Heavy), Iressa-Treated (Light)

TABLE: TYROSINE PHOSPHOSCAN® FINAL RESULTS, SILAC StuDY DEsiGN: Human non-small cell lung cancer (H3255) cell line; Trypsin Digest; Antibody: pY; CST #9411 Treatments: Untreated (Heavy), Iressa-treated (Light) Fold-Change Count (Iressa / in Index Untreated ) Protein Name Phosphorylation Site Description Peptide Accession kD Study Actin binding proteins 2 -1.6 CTNNA1 177 Catenin alpha-1 (Cadherin-associated protein) NAGNEQDLGIQyK P35221 100 4 3 -4.5 CTNND1 248 Catenin delta-1 (Cadherin-associated Src substrate) (p120(cas)) YRPSMEGyR O60716 108 2 4 -3.9 CTNND1 865 Catenin delta-1 (Cadherin-associated Src substrate) (p120(cas)) SQSSHSyDDSTLPLIDR O60716 108 1 5 -5.5 CTNND1 904 Catenin delta-1 (Cadherin-associated Src substrate) (p120(cas)) SLDNNySTPNERGDHNR O60716 108 1 6 -3 CTNND1 904 Catenin delta-1 (Cadherin-associated Src substrate) (p120(cas)) SLDNNySTPNER O60716 108 4 7 -1.8 CTNND1 §228 Catenin delta-1 (Cadherin-associated Src substrate) (p120(cas)) HYEDGYPGGSDNyGsLSR O60716 108 4 8 -1.8 CTNND1 §228 Catenin delta-1 (Cadherin-associated Src substrate) (p120(cas)) HYEDGYPGGSDNyGSLSR O60716 108 6 9 -1.1 CTNND1 §257 Catenin delta-1 (Cadherin-associated Src substrate) (p120(cas)) QDVyGPQPQVR O60716 108 1 0 -6. CTNND1 §280 Catenin delta-1 (Cadherin-associated Src substrate) (p120(cas)) FHPEPyGLEDDQR O60716 108 3 -3 CTNND1 §96 Catenin delta-1 (Cadherin-associated Src substrate) (p120(cas)) LNGPQDHSHLLySTIPR O60716 108 1 2 -. DBNL §162 Drebrin-like protein (SH3 domain-containing protein 7) (HIP-55) FQDVGPQAPVGSVyQK Q9UJU6 48 2 3 Adaptor/scaffold proteins 4 -0.7 ANK3 533 Ankyrin-3 (ANK-3) (Ankyrin-G) ADIVQQLLQQGASPNAATTSGyTPLHLSAR Q12955 480 3 5 -1.2 NEDD9 §166 Enhancer of filamentation 1 (CRK-associated substrate-related protein)(CasL) TGHGYVyEYPSR Q14511 93 4 6 -9.9 CRK 136 Proto-oncogene C-crk (p38) (Adapter molecule crk) QGSGVILRQEEAEyVR P46108 34 1 7 >-8.4 CRK 136 Proto-oncogene C-crk (p38) (Adapter molecule crk) QEEAEyVR P46108 34 1 8 >-58.4 DLG3 673 Disks large homolog 3 (Synapse-associated protein 102) (SAP102) RDNEVDGQDyHFVVSR Q92796 90 7 9 -25.5 DLG3 673 Disks large homolog 3 (Synapse-associated protein 102) (SAP102) DNEVDGQDyHFVVSR Q92796 90 3 20 -9.9 EPS8 §485 Epidermal growth factor receptor kinase substrate 8 LSTEHSSVSEyHPADGYAFSSNIYTR Q12929 92 2 2 -1.3 FLOT1 203 Flotillin-1 VSAQyLSEIEMAK O75955 47 3 22 -3.9 GAB1 §406 GRB2-associated-binding protein 1 (GRB2-associated binder 1) DASSQDCyDIPR Q13480 77 2 23 >-.0 GAB1 §627 GRB2-associated-binding protein 1 (GRB2-associated binder 1) GDKQVEyLDLDLDSGK Q13480 77 4 24 >-22.0 GAB1 §659 GRB2-associated-binding protein 1 (GRB2-associated binder 1) SSGSGSSVADERVDyVVVDQQK Q13480 77 3 25 -8.7 ITSN2 §552 Intersectin-2 (SH3 domain-containing protein 1B) (SH3P18) LIyLVPEK Q9NZM3 193 2 26 -1 WASL §256 Neural Wiskott-Aldrich syndrome protein (N-WASP) VIyDFIEK O00401 55 2 27 -1.6 PAG1 §227 Phosphoprotein associated with glycosphingolipid-enriched microdomains 1 AEFAEyASVDR Q9NWQ8 47 4 28 -3.2 PAG1 §341 Phosphoprotein associated with glycosphingolipid-enriched microdomains 2 SGQSLTVPESTyTSIQGDPQR Q9NWQ8 47 1 29 -2.6 PAG1 §359 Phosphoprotein associated with glycosphingolipid-enriched microdomains 3 SPSSCNDLyATVK Q9NWQ8 47 4 30 >-0.4 PAG1 §417 Phosphoprotein associated with glycosphingolipid-enriched microdomains 4 ATLGTNGHHGLVPKENDyESISDLQQGR Q9NWQ8 47 4 3 -3.2 PAG1 §417 Phosphoprotein associated with glycosphingolipid-enriched microdomains 5 ENDyESISDLQQGR Q9NWQ8 47 4 32 -1.7 PARD3 388 Partitioning-defective 3 homolog (Atypical PKC isotype-specific-interacting prot) FSPDSQyIDNR Q8TEW0 151 2 33 -1.3 PARD3 489 Partitioning-defective 3 homolog (Atypical PKC isotype-specific-interacting prot) DVTIGGSAPIyVK Q8TEW0 151 4 34 -1.4 PARD3 719 Partitioning-defective 3 homolog (Atypical PKC isotype-specific-interacting prot) RISHSLySGIEGLDESPSR Q8TEW0 151 1 35 -0.9 PARD3 719 Partitioning-defective 3 homolog (Atypical PKC isotype-specific-interacting prot) ISHSLySGIEGLDESPSR Q8TEW0 151 5 36 -0.3 PARD3 719 Partitioning-defective 3 homolog (Atypical PKC isotype-specific-interacting prot) RIsHSLySGIEGLDESPSR Q8TEW0 151 2 37 -0.8 PARD3 §1127 Partitioning-defective 3 homolog (Atypical PKC isotype-specific-interacting prot) EGHMMDALyAQVK Q8TEW0 151 1 38 -2.1 MPZL1 §263 Myelin protein zero-like protein 1 SESVVyADIR O95297 29 3 39 -1.5 DLG1 760 Disks large homolog 1 (Synapse-associated protein 97) (SAP-97) (hDlg) DyHFVTSR Q12959 100 1 40 -1.2 DLG1 760 Disks large homolog 1 (Synapse-associated protein 97) (SAP-97) (hDlg) DYEVDGRDyHFVTSR Q12959 100 3 4 -1.1 DLG1 760 Disks large homolog 1 (Synapse-associated protein 97) (SAP-97) (hDlg) RDYEVDGRDyHFVTSR Q12959 100 1 42 -1 SHB §201 SH2 domain-containing adapter protein B LDyCGGSGEPGGVQR Q15464 55 2 43 -1.3 SHB §333 SH2 domain-containing adapter protein B DKVTIADDySDPFDAK Q15464 55 4 44 -1.1 SHB §333 SH2 domain-containing adapter protein B DKVTIADDySDPFDAKNDLK Q15464 55 5 45 -1.1 SHB 355 SH2 domain-containing adapter protein B AGKGESAGyMEPYEAQR Q15464 55 6 46 -1.1 SHB 355 SH2 domain-containing adapter protein B GESAGyMEPYEAQR Q15464 55 3 47 >-8. SHC1 §427 SHC-transforming protein 1 (SH2 domain protein C1) ELFDDPSyVNVQNLDK P29353 63 7 LEGEND: § = published site, * - phosphorylation, # = oxidized methionine SUMMARY TABLE TABLE: TYROSINE PHOSPHOSCAN® FINAL RESULTS, SILAC StuDY DEsiGN: Human non-small cell lung cancer (H3255) cell line; Trypsin Digest; Antibody: pY; CST #9411 Treatments: Untreated (Heavy), Iressa-treated (Light) Fold-Change Count (Iressa / in Index Untreated ) Protein Name Phosphorylation Site Description Peptide Accession kD Study 48 >-3.9 SPRY1 53 Sprouty homolog 1 GSNEyTEGPSVVK O43609 35 1 49 >-. SPRY1 89 Sprouty homolog 1 THEIIPINVNNNyEHR O43609 35 2 50 >-5.8 SPRY4 §75 Sprouty homolog 4 TSHVENDyIDNPSLALTTGPKR Q6QIX2 35 1 5 >-.3 TNS3 §780 Tumor endothelial marker 6 KLsLGQyDNDAGGQLPFSK Q8IZW7 155 2 52 -8.7 TNS3 §780 Tumor endothelial marker 6 LSLGQyDNDAGGQLPFSK Q8IZW7 155 1 53 -7.4 TNS3 §780 Tumor endothelial marker 6 KLSLGQyDNDAGGQLPFSK Q8IZW7 155 5 54 -1.1 TNS1 1404 Tensin-1 AGSLPNyATINGK Q9HBL0 186 2 55 -1.9 TJP2 §1118 Tight junction protein ZO-2 (Zonula occludens protein 2) IEIAQKHPDIyAVPIK Q9UDY2 134 3 56 Adhesion proteins 57 >-.8 CTNNB1 30 Catenin beta-1 AAVSHWQQQSyLDSGIHSGATTTAPSLSGK P35222 86 2 58 -2.6 CDH1 §753 Epithelial cadherin DNVyYYDEEGGGEEDQDFDLSQLHR P12830 98 2 59 -2.1 ERBB2IP §1104 Protein LAP2 (Erbb2-interacting protein) (Erbin) (Densin-180-like protein) AQIPEGDyLSYR Q96RT1 158 6 60 -3. ERBB2IP §1104 Protein LAP2 (Erbb2-interacting protein) (Erbin) (Densin-180-like protein) RAQIPEGDyLSYR Q96RT1 158 8 6 -7.8 ERBB2IP 1164 Protein LAP2 (Erbb2-interacting protein) (Erbin) (Densin-180-like protein) TMSVSDFNySR Q96RT1 158 2 62 -3 F11R 280 Junctional adhesion molecule A KVIySQPSAR Q9Y624 33 4 63 -2.1 F11R 280 Junctional adhesion molecule A VIySQPSAR Q9Y624 33 2 64 -33.7 ITGB1 §783 Integrin beta-1 WDTGENPIyKSAVTTVVNPK P05556 89 1 65 -2.4 ITGB1 §783 Integrin beta-1 WDTGENPIyK P05556 89 4 66 -03.9 ITGB1 §795 Integrin beta-1 SAVTTVVNPKyEGK P05556 89 1 67 -1.7 MUC1 §1203 Mucin-1 DTyHPMSEYPTYHTHGR P15941 122 2 68 -0.8 MUC1 1209 Mucin-1 DTYHPMSEyPtYHTHGR P15941 122 1 69 -0.9 MUC1 1209, §1212 Mucin-1 DTYHPMSEyPTyHTHGR P15941 122 1 70 -1.6 MUC1 §1212 Mucin-1 DTYHPMSEYPTyHTHGR P15941 122 2 7 -1.9 MUC1 §1229 Mucin-1 YVPPSSTDRSPyEK P15941 122 3 72 -1.8 PVRL1 468 Poliovirus receptor-related protein 1 YDEDAKRPyFTVDEAEAR Q15223 57 4 73 -0.7 PVRL4 445 Poliovirus receptor-related protein 4 SySTLTTVR Q96K15 55 1 74 -0.9 PVRL4 502 Poliovirus receptor-related protein 4 AKPTGNGIyINGR Q96K15 55 4 75 >-7.9 OCLN 287 Occludin SNILWDKEHIyDEQPPNVEEWVK Q16625 59 1 76 -6.3 PKP2 166 Plakophilin-2 AHYTHSDyQYSQR Q99959 97 4 77 -5. PKP2 631 Plakophilin-2 YSQNIyIQNR Q99959 97 1 78 -6.8 PKP3 §84 Plakophilin-3 GQyHTLQAGFSSR Q9Y446 87 9 79 -2.7 PKP3 176 Plakophilin-3 ADyDTLSLR Q9Y446 87 7 80 -1 PKP3 176 Plakophilin-3 GGVGSRADyDTLSLR Q9Y446 87 2 8 -1.1 PKP3 390 Plakophilin-3 NLIyDNADNK Q9Y446 87 1 82 -4.5 PKP4 157 Plakophilin-4 (p0071) SSTQMNSYSDSGyQEAGSFHNSQNVSK Q99569 134 1 83 -0.8 PKP4 372 Plakophilin-4 (p0071) TVHDMEQFGQQQYDIyER Q99569 134 4 84 -0.7 PKP4 §415 Plakophilin-4 (p0071) SAVSPDLHITPIyEGR Q99569 134 3 85 -1.6 PKP4 470 Plakophilin-4 (p0071) NNyALNTTAtYAEPYRPIQYR Q99569 134 1 86 -1.5 PKP4 470, §478 Plakophilin-4 (p0071) NNyALNTTATyAEPYRPIQYR Q99569 134 2 87 -1.1 PKP4 §478 Plakophilin-4 (p0071) NNYALNTTATyAEPYRPIQYR Q99569 134 2 88 -1.3 PKP4 1168 Plakophilin-4 (p0071) STTNyVDFYSTK Q99569 134 7 89 -1.7 SDC4 197 Syndecan-4 KAPTNEFyA P31431 22 2 90 -0.7 VCL §821 Vinculin (Metavinculin) SFLDSGyR P18206 124 4 9 Cytoskeletal proteins 92 -1 KRT7 39 Keratin, type II cytoskeletal 7 (Cytokeratin-7) (Sarcolectin) LSSARPGGLGSSsLyGLGASRPR P08729 51 2 93 -0.7 KRT7 39 Keratin, type II cytoskeletal 7 (Cytokeratin-7) (Sarcolectin) LSSARPGGLGSSSLyGLGASRPR P08729 51 4 94 -8.4 CLDN3 214 Claudin-3 (CPE-receptor 2) (HRVP1) STGPGASLGTGyDR O15551 23 1 2 LEGEND: § = published site, * - phosphorylation, # = oxidized methionine SUMMARY TABLE TABLE: TYROSINE PHOSPHOSCAN® FINAL RESULTS, SILAC StuDY DEsiGN: Human non-small cell lung cancer (H3255) cell line; Trypsin Digest; Antibody: pY; CST #9411 Treatments: Untreated (Heavy), Iressa-treated (Light) Fold-Change Count (Iressa / in Index Untreated ) Protein Name Phosphorylation Site Description Peptide Accession kD Study 95 -9.3 CLDN3 219 Claudin-3 (CPE-receptor 2) (HRVP1) STGPGASLGTGYDRKDyV O15551 23 3 96 -5.8 CFL1 §139 Cofilin-1 (Cofilin, non-muscle) LTGIKHELQANCyEEVKDR P23528 19 2 97 -4.3 CFL1 §139 Cofilin-1 (Cofilin, non-muscle) HELQANCyEEVKDR

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