Fig. 1

Proteomic analysis of RCC secretome. A Left: The scheme of the experiment. CM from all cell lines was subjected to proteomic and metabolomics analysis. Cells were collected and used for transcriptomic analysis. Right: nanoHPLC-MS/MS revealed 85 CM proteins that were consistently altered in all ccRCC cell lines the same direction (upregulated/downregulated) and verified as secretome components. Proteins selected for validation are shown in red font. Complete data are shown in Additional file 1: Table S4. The analysis was performed using protein extracts isolated from 3 independent biological experiments per each cell line. B Gene ontology analysis of the 85 consistently altered proteins. C Validation of top altered proteins using ELISA. The analysis was performed using conditioned media isolated from 3 independent biological experiments per each cell line. Statistical analysis: One-way ANOVA with Dunnett's Multiple Comparison Test. *p < 0.05, **p < 0.01, ***p < 0.001. D The expression of genes encoding altered CM proteins is disturbed in ccRCC cell lines. The table shows result of microarray analysis (complete data are shown in Additional file 1: Table S5). The plots show results of qPCR validation of the analyzed genes in RNA isolated from 3 independent biological experiments. Statistical analysis: One-way ANOVA with Dunnett's Multiple Comparison Test. *p < 0.05, **p < 0.01, ***p < 0.001