Fig 1: Resident memory T (TRM) and non-TRM phenotypic profiles and susceptibility of cancer stem cell (CSC) to cytotoxic T lymphocyte (CTL)-mediated killing. (A) Flow cytometry analyses of CD8, LFA-1, CD103, CD49a, CD69, PD-1, CTLA-4, TGFBR2 and VEGFR2 on CD8+CD103+ TRM (Heu171) and CD8+CD103- non-TRM (H32-22) clones. Mean immunofluorescence intensity (MFI) are in parentheses.(B) Cytotoxic activity of the TRM clone (Heu171) towards autologous IGR-Heu, Heu-CSC, CSC-1 and CSC-2 target cells. Percent of specific lysis are shown at indicated effector to target (E:T) ratios. (C) Quantification of transforming growth factor (TGF)-ß in conditioned media from IGR-Heu, Heu-CSC, CSC-1 and CSC-2 by multi-analyte flow assay. Ratios of active/total TGF-ß normalized to IGR-Heu are included. Results are presented as mean±SEM of duplicates. (D) Cytotoxicity of the non-TRM clone (H32-22) towards autologous IGR-Heu, Heu-CSC, CSC-1 and CSC-2 target cells. (E) Inhibition of TRM-cell-mediated killing. CSC-1 cells are preincubated in the presence of isotype control, anti-major histocompatibility complex class I (MHC-I) or anti-intercellular adhesion molecule 1 (ICAM-1) neutralizing monoclonal antibody (mAb) and then CTL were added at 5:1 E:T ratio. Symbols represent replicates and horizontal bars represent means±SEM (n=3). P value was determined by two-way analysis of variance test. *p<0.05, **p<0.01,***p<0.001,****p<0.0001. Data are from one representative experiment out of three.