Journal: Journal for Immunotherapy of Cancer
Article Title: Cancer stem-like cells evade CD8 + CD103 + tumor-resident memory T (T RM ) lymphocytes by initiating an epithelial-to-mesenchymal transition program in a human lung tumor model
doi: 10.1136/jitc-2022-004527
Figure Lengend Snippet: Phenotypic and molecular characterization of lung cancer stem cells (CSC). (A) Flow cytometry profiles of CD34, CD44, PECAM-1 (CD31), CD24, CD166, C-X-C chemokine receptor type 4 (CXCR4), vascular endothelial growth factor (VEGF) and epidermal growth factor receptor (EGFR) in IGR-Heu, Heu-CSC, CSC-1 and CSC-2. Percentages of positive cells and mean immunofluorescence intensity (MFI) (in parentheses) are shown. (B) Western blot analysis of NANOG, SNAIL1 and SOX2 proteins in IGR-Heu and CSC. Right, normalization of proteins relative to F-actin. (C) Western blot analysis of total GSK-3β and phospho-GSK-3β proteins in IGR-Heu, Heu-CSC, CSC-1 and CSC-2. Right, ratio of phospho-GSK-3β to total GSK-3β. Data are from one representative experiment out of three. (D) Flow cytometry profiles of major histocompatibility complex class I (MHC-I) and HLA-A2 on IGR-Heu, Heu-CSC, CSC-1 and CSC-2. Percentage of positive cells and MFI (in parentheses) are included. (E) Flow cytometry analyses of E-cadherin and intercellular adhesion molecule (ICAM)-1 expression on IGR-Heu, Heu-CSC, CSC-1 and CSC-2. Percentage of positive cells and MFI are shown.
Article Snippet: For tumor cell and CSC phenotyping, anti-CD24 (SN3, Invitrogen MHCD2404, 1/100), anti-CD34 (AC136, Miltenyi 130-113-182, 1/100), anti-CD44 (DB105, Miltenyi 130-113-896, 1/100), anti-CD166 (3A6, BD 562131, 1/100), anti-CXCR4 (12G5, Biolegend 306510, 1/100), anti-VEGF (23410, R&D IC2931A, 1/100), anti-epidermal growth factor receptor (EGFR) (AY13, Biolegend 352913, 1/100), anti-major histocompatibility complex class I (MHC-I) (W6/32, Biolegend 311429, 1/100), anti-HLA-A2 (BB7.2, BD 558570, 1/100), anti-programmed death-ligand 1 (PD-L1) (29E.2A3, Biolegend 329721, 1/100), anti-programmed death-ligand 2 (PD-L2) (MIH18, Biolegend 345515, 1/100), anti-B7-1 (2D10, Biolegend 305207, 1/100), anti-B7-2 (BU63, Biolegend 374215, 1/100), anti-E-cadherin (67A4, Biolegend 324114, 1/100) and anti-ICAM-1 (HA58, Biolegend 353113, 1/100) monoclonal antibodies (mAb) were used.
Techniques: Flow Cytometry, Immunofluorescence, Western Blot, Expressing
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