Fig. 1: Purification of pericytes from normal adjacent tissues and tumors derived from NSCLC and HCC patients using CD146^+--FITC-MACS enrichment method.

a–f Representative triple immunostaining images of CD34 (white), CD146 (green), and α-SMA (red) from 2 different microscopic fields in normal adjacent tissues and tumors derived from NSCLC and HCC patients are given. Bar charts represent the percentage of CD146 and α-SMA-double positive (+ve) blood vessels per field in each group (n = 10 NSCLC/HCC patients). Statistical tests were two-sided. g–j Representative FACS plots showing the gating strategy for the purification of pericytes from normal adjacent tissues and tumors derived from NSCLC patients with (+) or without (−) CD146⁺-FITC-microbead cell soring enrichment (MACS) method. Debris, doublets (TOF), and dead cells (FVD) were excluded, then CD45⁻CD31⁻CD146⁺PDGFRβ⁺ pericytes were collected. Percentage refers to the proportion of cells in the previous parent gate. k, l Bar charts show the relative fold change in percentage of pericytes and endothelial cells isolated by the MACS enrichment coupled with FACS protocol compared to that isolated by FACS alone. Statistical tests were two-sided. Percentages of pericyte refer to cell population in the R1 gate. NSCLC non-small cell lung cancer, HCC hepatocellular carcinomas, FSC forward-scatter; SSC side-scatter, TOF time of fly, FVD fixable viability dye, EC endothelial cells, PC pericytes, NEC normal adjacent tissue derived endothelial cells, TEC tumor-derived endothelial cells, NPC normal adjacent tissue derived pericytes, TPC tumor-derived pericytes. FACS plots/bar charts are representative of three individual patient data. Means ± SEM are given. c, f, k, l Student’s t test. Scale bars in a, b, d, e represent 50 μm, a, b, d, e (magnified pictures) represent 25 μm.