波形蛋白在肿瘤内皮细胞中过表达,细胞外存在。A,B波形蛋白mRNA(a; n = 3; qPCR)和蛋白质(B; n = 4;流式细胞仪)在人类结肠肿瘤(TEC)和正常结肠(NEC)中分离的内皮细胞(EC)中的表达。C中的c波形蛋白mRNA表达在人(结肠,n = 13;神经胶质瘤,n = 2)和鼠(神经胶质瘤,n = 7;黑色素瘤,n = 2)肿瘤中分离的EC中。D蛋白质组学分析分泌组中的人类正常结肠和结直肠癌组织(左图; n = 21(正常),n = 4(I级),n = 8(II级),n = 5iv))和总细胞内波形蛋白(右图; n = 15(正常),n = 15(CRC I – IV))。数据表示为A – D中的平均值±SEM。p值代表配对的t检验(A,C,D右图),未配对的t检验(B)和单向ANOVA(D左图)。固定和通透的HUVEC(左图)和活的完整HUVEC(右图)的E免疫荧光染色。插图:负面对照。显示了至少三个独立实验的代表性图像。 f Schematic representation of vimentin localization (in green). g Western blotting of total cell lysate, ECM deposit, and secretome of HUVEC. Representative sections of at least three independent experiments are shown. h Global proteomics analysis (n = 1) of HUVEC lysate, secretome, and ECM deposit. i (Left) Proportion of known tumor EC markers (TEC, red) among externalized proteins. (Right) Secretion mechanisms among externalized proteins. j Protein–protein interaction analysis using STRING of externalized TEC markers. Opacity levels of the nodes are proportional to secretion abundance. k Effect of angiogenesis inhibitors and cytokines on vimentin secretion. Relative secretion is color-coded according to the legend right of the panel, and agent types are color-coded according to the legend below the panel. l Schematic of different cellular protein secretion pathways. m Effect of different protein secretion mediators on vimentin secretion. Legend as in k. Data are color-coded as mean values of relative secretion in k and m; numbers of samples are presented in the Source Data file. *p ﹤0.05 based on Kruskal–Wallis test with Dunn’s multiple comparison test correction for k and m. Source data are provided as a Source Data file. Credit:自然通讯(2022)。doi:10.1038/s41467-022-30063-7