Verified MultiOmyx™ Multiplexing Panels Immune Panel (TIL panel) – 12 markers Tested in multiple cancer indications (melanoma, lung, colorectal, prostate, breast) TIL Panel Co-expression Phenotypes CD3 CD3+CD4+ T helper CD4 CD3+CD4+FoxP3+ T regulatory CD8 CD3+CD4+CD45RO+ Memory T helper CD45RO CD3+CD4+PD1+ Immune modulation FoxP3 CD3+CD8+ T cytotoxic CD20 CD3+CD8+CD45RO+ Memory T cytotoxic CD68 CD3+CD8+PD1+ Immune modulation CD56 CD68+ Macrophage PanCK+ CD8+ PD-L1+ CD68+ FoxP3+ Au Q. et al. (April, 2016). MultiOmyx™ multiplexed tumor infiltrating lymphocyte CTLA-4 CD68+PDL1+ Macrophage PD-L1 panel provides comprehensive immunophenotyping from a single FFPE slide. Poster presented at AACR Annual Meeting, New Orleans, LA. PD-1 CD20+ B cell PD-L1 CD20+PDL1+ B cell PD-L1 PanCK CD3-CD56+ Natural Killer cell PanCK+PDL1+ Tumor cell PD-L1 Myeloid Panel – 10 markers Tested in PDAC tissue Myeloid Panel Co-expression Phenotypes CD11b CD11b+CD33+ Myeloid cells M1 TAM CD14 CD11b+CD33+HLADR- MDSC CD15 CD11b+CD33+HLADR-CD14+CD15- M-MDSC CD16 CD11b+CD33+HLADR-CD14-CD15+ G-MDSC CD33 CD68+ TAM CD68 CD68+HLADR+CD163- M1 TAM CD163 CD68+HLADR-CD163+ M2 TAM HLA-DR CD11b+HLADR-CD16+ Neutrophil/G-MDSC PanCK+ CD68+ CD11b+ HLA-DR+ Juncker-Jensen A. et al. (April, 2018). Tumor-Infiltrating Myeloid Cells – Using Arginase1 MultiOmyx™ to Distinguish between MDSCs, TAMs and TANs in the Pancreatic Tumor Microenvironment. Poster presented at AACR Annual Meeting, Chicago, IL. PanCK Verified MultiOmyx™ Multiplexing Panels TIL & Myeloid Panel #1 – 16 markers Tested in NSCLC tissue TIL/TAM Panel #1 Co-expression Phenotypes CD3 CD3+CD4+ T helper LAG3+/PD1+ CTL CD4 CD3+CD4+FoxP3+ T regulatory CD8 CD3+CD4+PD1+ Immune modulation FoxP3 CD3+CD8+ T cytotoxic CD20 CD8+PD1+ Immune modulation CD68 CD3+TIM3+ Immune modulation CD163 CD3+LAG3+ Immune modulation CD11b CD3+ICOS+ Immune modulation PanCK+ LAG-3+ PD-1+ CD8+ Juncker-Jensen A. et al. (2019, December). PD-1 and LAG-3 synergize to drive CD15 CD3+OX40+ Immune modulation tumor-infiltration of T cytotoxic cells in NSCLC tumors. Poster presented at ESMO I/O Annual Meeting, Geneva, Switzerland. TIM-3 CD20+ B cell LAG-3 CD11b+CD15+ Granulocyte OX40 CD68+ TAM ICOS CD68+CD163+ M2 TAM PD-1 CD68+PDL1+ Macrophage PD-L1 PD-L1 CD68+TIM3+ Macrophage TIM-3 PanCK PanCK+PDL1+ Tumor cell PD-L1 TIL & Myeloid Panel #2 – 19 markers Tested in PDAC tissue TIL/Myeloid Panel Co-expression Phenotypes #2 CD3+CD4+ T helper CD3 CD3+CD4+FoxP3+ T regulatory G-MDSC CD4 CD3+CD4+PD1+ Immune modulation CD8 CD3+CD8+ T cytotoxic CD45RO CD8+PD1+ Immune modulation FoxP3 CD8+GranzymeB+ Effector T cytotoxic CD11b CD3+CD45RO Memory T cells CD14 CD68+ TAM PanCK+ CD11b+ CD3+ CD15+ FoxP3+ ™ CD15 Juncker-Jensen A. et al. (October, 2018). Using MultiOmyx to Analyze CD68+HLADR+CD163- M1 TAM Correlations between Immunosuppressive Cells and Tumor-Infiltrating CD16 Lymphocytes in the Pancreatic Tumor Microenvironment. Poster presented at CD68+HLADR-CD163+ M2 TAM ESMO Annual Meeting, Munich, Germany. CD33 CD68+PDL1+ Macrophage PD-L1 CD68 CD11b+CD33+ Myeloid cells CD163 CD11b+CD33+HLADR- MDSC HLA-DR CD11b+CD33+HLADR-CD14+CD15- M-MDSC Arginase1 CD11b+CD33+HLADR-CD14-CD15+ G-MDSC GranzymeB PanCK+Ki67+ Proliferating tumor Ki67 PanCK+PDL1+ Tumor cell PD-L1 PD-1 PD-L1 PanCK 2 | Verified MultiOmyx™ Multiplexing Panels Dendritic Cell Panel – 15 markers Tested in melanoma tissue Dendritic Panel Co-expression Phenotypes CD3 CD3+CD4+ T helper pDC CD4 CD3+CD8+ T cytotoxic CD8 CD68+ TAM CD11c CD68+HLADR+CD163- M1 TAM CD14 CD68+HLADR-CD163+ M2 TAM CD40 CD123+HLADR+ pDC CD68 CD123+HLADR+CD40+ CD40 positive pDC CD123 CD123+HLADR+DCLAMP+ Activated pDC CD141 CD11c+Clec9A+CD141+HLADR+ cDC CD163 CD11c+Clec9A+CD141+CD40+ CD40 positive cDC SOX10+CD123+HLADR+CD11c+ Gozo M. et al. (2020, June). Distinguishing dendritic cell subtypes in the HLA-DR CD11c+Clec9A+CD141+DCLAMP+ Activated cDC tumor microenvironment using MultiOmyxTM. Poster presented at AACR Annual Meeting, Virtual. Clec9A CD11c+CD14+DCSIGN+HLADR+ Monocyte derived DC DC-SIGN DC-LAMP SOX10/PanCK Tertiary Lymphoid Structure (TLS) Panel – 14 markers Tested in bladder cancer tissue TLS Panel Co-expression Phenotypes CD3 CD3+CD4+ T helper CD4 CD3+CD4+FoxP3+ T regulatory CD8 CD3+CD8+ T cytotoxic FoxP3 CD20+ B cell CD11b CD11b+HLADR+ APC myeloid cell CD14 CD11b+HLADR- MDSC CD15 CD11b+HLADR-CXCR2+ MDSC positive for CXCR2 CD20 CD11b+HLADR-iNOS MDSC positive for iNOS CD68 CD11b+HLADR-CD14+CD15- M-MDSC HEV PNAd CD11b+HLADR-CD14+CD15- G-MDSC CXCR2 CD68+ TAM iNOS CD68+HLADR+ M1 TAM CD20+ PNAd+ CD3+ Image of TLS in a bladder cancer sample from an unpublished academic HLA-DR CD68+iNOS+ M1 TAM collaboration study with University of Notre Dame. PanCK PNAd+ High Endothelial Venule PanCK+iNOS+ iNOS positive tumor cells Verified MultiOmyx™ Multiplexing Panels | 3 TIL, TAM & Vessel Panel #1 – 10 markers Tested in breast cancer tissue TIL/TAM Panel #1 Co-expression Phenotypes CD3 CD3+CD4+ T helper CD4 CD3+CD4+FoxP3+ T regulatory CD8 CD3+CD8+ T cytotoxic FoxP3 CD68+ TAM CD68 CD68+HLADR+CD163- M1 TAM CD163 CD68+HLADR-CD163+ M2 TAM PanCK+ CD3+ CD34+ Ki67+ HLA-DR CD34+ Vessels Juncker-Jensen A. et al. (April, 2019). Pro-Tumorigenic Mechanisms of M2 Tumor- Associated Macrophages in Triple-Negative Breast Cancer. Poster presented at Ki67 PanCK+Ki67+ Proliferating tumor AACR Annual Meeting, Chicago, IL. CD34 PanCK TIL, TAM & Vessel Panel #2 – 16 markers Tested in ovarian tumor tissue TIL/TAM Panel #2 Co-expression Phenotypes M1 TAM CD3 CD3+CD4+ T helper CD4 CD3+CD4+FoxP3+ T regulatory M2 TAM CD8 CD3+CD4+PD1+ Immune modulation FoxP3 CD4+CTLA4+ Immune modulation CD20 CD3+CD8+ T cytotoxic CD68 CD8+PD1+ Immune modulation CD163 CD8+CTLA4+ Immune modulation HLA-DR CD20+ B cell S100+ CD68+ HLA-DR+ CD163+ PD-1 CD68+ TAM Juncker-Jensen A. et al. (2019, October). An Integrated Multiplexing Approach for the Immunoprofiling of the Tumor Microenvironment of Ovarian Granulosa Cell PD-L1 CD68+HLADR+CD163- M1 TAM Tumors. Poster presented at AACR-NCI-EORTC Annual Meeting, Boston, MA. CTLA-4 CD68+HLADR-CD163+ M2 TAM CD34 CD68+PDL1+ Macrophage PD-L1 Vimentin CD34+ Vessels S100 PanCK+Ki67+ Proliferating tumor Ki67 PanCK+PDL1+ Tumor cell PD-L1 PanCK 4 | Verified MultiOmyx™ Multiplexing Panels TIGIT Immune Cell Panel – 15 markers Tested in NSCLC & melanoma tissue TIGIT Immune Panel Co-expression Phenotypes TIGIT CD3+CD4+ T helper CD226 CD3+CD4+FoxP3+ T regulatory CD155 CD3+CD4+PD1+ Immune modulation CD3 CD3+CD8+ T cytotoxic CD4 CD8+PD1+ Immune modulation CD8 CD8+GranzymeB+ Effector T cytotoxic FoxP3 CD68+ TAM SOX10 + TIGIT+ LAG3+ PD1+ Au Q. et al. (April, 2019). Characterization of TIGIT Expression Using MultiOmyx™ Hyperplexed Immunofluorescence Assay in NSCLC and melanoma. CD56 CD68+HLADR+CD163- M1 TAM Poster presented at AACR Annual Meeting, Atlanta, GA. CD45RO CD68+HLADR-CD163+ M2 TAM CD11b CD68+PDL1+ Macrophage PD-L1 CD11c CD11b+CD33+ Myeloid cells PD-1 CD11b+CD33+HLADR- MDSC LAG-3 CD11b+CD33+HLADR-CD14+CD15- M-MDSC TIM-3 CD11b+CD33+HLADR-CD14-CD15+ G-MDSC PanCK/SOX10 PanCK+Ki67+ Proliferating tumor PanCK+PD1+ Tumor cell PD-L1 Myeloid Leukemia Immune Panel – 14 markers Tested in AML/HL/DLBCL Leukemia Panel Co-expression Phenotypes CD3 CD3+CD4+ T helper G-MDSC CD4 CD3+CD4+FoxP3+ T regulatory CD8 CD3+CD8+ T cytotoxic G-MDSC FoxP3 CD68+ TAM G-MDSC CD68 CD68+HLADR+CD163- M1 TAM CD163 CD68+HLADR-CD163+ M2 TAM HLA-DR CD11b+CD33+ Myeloid cells CD11b CD11b+CD33+HLADR- MDSC CD11b+CD33+CD15+CD34+ Au Q. et al. (2019, December). Phenotypic Characterization of the Immune CD33 CD11b+CD33+HLADR-CD14+CD15- M-MDSC Landscape in the Bone Marrow of Patients with Acute Myeloid Leukemia (AML) Using MultiOmyx™ Hyperplexed Immunofluorescence CD14 CD11b+CD33+HLADR-CD14-CD15+ G-MDSC Assay. Poster presented at ASH Annual Meeting, Orlando, FL. CD15 CD11b+CD33+HLADR-CD14-CD15+Arg1 Arg1+ G-MDSC CD34 Arginase1 Tumor marker Verified MultiOmyx™ Multiplexing Panels | 5 Hepatitis B (HBV) Panel – 19 markers Tested in liver tissue HBV Panel Co-expression Phenotypes HBcAG CD3+CD4+ T helper HBsAG CD3+CD4+FoxP3+ T regulatory NaKATPase CD3+CD4+PD1+ Immune modulation CD3 CD3+CD8+ T cytotoxic CD4 CD3+CD8+PD1+ Immune modulation CD8 CD68+ TAM FoxP3 CD68+HLADR+ M1 macrophage CD56 CD68+CD163+ M2 macrophage HBcAG+ HBsAG+ DAPI+ CD20 CD68+PDL1+ Macrophage PD-L1 CD11b CD68+DCSIGN+ Macrophage DC-SIGN HLA-DR CD11b+HLADR- MDSC CD11c CD11c+HLADR+ Dendritic cell (cDC) DC-SIGN CD11c+HLADR+DCSIGN+ Dendritic cell CD68 CD20+ B cell Dendritic cell CD163 CD20+HLADR+ B cell HLA-DR PD-1 CD56+CD3- NK cell PD-L1 CD56+CD3-HLADR+ NK cell HLA-DR CD11c+ HLADR+ CD68+ CD20+ IDO1 Ki67 NASH Liver Panel – 17 markers Tested in liver tissue NASH Panel Co-expression Phenotypes CD11b CD11b+CD11c+CD14-HLADR+ cDC HLA-DR CD11c+HLADR+CD14-IDO1+ cDC IDO1+ CD68 CD11b+CD11c+HLADR+CD14-Ki67+ cDC proliferating CD163 CD68+CD163+CD16 Kupffer cell Kupffer cell CD11c CD68+CD163+CD16+DCSIGN+ Kupffer cell DC-SIGN+ DC-SIGN CD68+CD163+CD16+PDL1+ Kupffer cell PD-L1+ CD66b Kupffer cell prolifer- CD68+CD163+CD16+Ki67+ ating CD9 CD68+ CD16+ CD163+ CD68+CD163+CD16+CD33+ Kupffer cell CD33+ CD14 CD11b+CD14+HLADR+ Tmo CD16 CD11b+CD14+HLADR+CD16+ TMo CD16+ CD33 HLADR+CD9+CD14+ SAM SMA HLADR+CD9+CD14+TREM2+ SAM TREM2+ TREM2 HLADR+CD9+CD14+PDL1+ SAM PD-L1+ MNDA HLADR+CD9+CD14+DCSIGN+ SAM DC-SIGN+ IDO1 HLADR+CD9+CD14+CD33+ SAM CD33+ PD-L1 HLADR+CD9+CD14+MNDA+ SAM MNDA+ Ki67 HLADR+ CD16+ CD14+ CD9+ HLADR+CD9+CD14+Ki67+ SAM proliferating PD-L1 PanCK 6 | Verified MultiOmyx™ Multiplexing Panels Verified MultiOmyx™ Multiplexing Panels | 7 About NeoGenomics NeoGenomics is an industry-leading cancer diagnostics and pharma services company.