Fig 1: Isolated Lin-EpCAM-CD73+CD90+ mesenchymal cell subset display a perivascular-like phenotype. (A) Representative bivariate flow cytometric plots showing the expression of PDGFR-beta PDGFR-alpha in Lin-EpCAM-CD73+CD90+ cells isolated from tumor and matched normal (upper panels) specimens, as well as human lung fibroblasts (HLFib, CCD-16Lu, ATCC® CCL-204™) and BM-MSC (lower panels). (B) Expression of CD105 (left panels) and NG2 (right panels) for the subsets of cells in gate R1 and R2 demonstrated in histogram overlay. (C) Scatter plots showing the MFI of CD105 and NG2 in the cell subsets R1 and R2 (n = 10, biological replicates, see Figure S5 for FMO controls for setting cell boundaries). (D) mRNA expression of selected genes of various mesenchymal markers and functional categories specific to the lung in sorted Lin-EpCAM-CD73+CD90+ (n = 8, biological replicates). HLFib was set at one. (E) Representative histogram overlay showing the change in expression of PD-L1 in normal and tumor-derived Lin-EpCAM-CD73+CD90+ cells (upper panels) in response to exposure to proinflammatory cytokines, as well as in HLFib and BM-MSC (lower panels). (F) Bar graphs showing the change in MFI for PD-L1 (n = 8, biological replicates). Data in (D and F) are presented as mean ± SD. Error bars show SD. Statistical analysis in C by Student t-test, two-tailed, for comparison of paired or unpaired parametric data. All tests were two-tailed. Statistical analysis of means for more than two groups in (D and F) were by one-way ANOVA and multiple comparisons using post hoc Newman-Keuls test. *p < 0.05 were considered significant. See related supplementary data Figures S5–S7.