Parallel mRNA, proteomics and miRNA expression analysis in cell line models of the intestine

Figure 1 Overview of Tri-Omics data analysis approach. Stage 1: Differential expression analysis of “Tri-Omics” data (miRNA, protein, mRNA) between HT-29 and Caco-2 cell lines (for a full list of results, including fold-changes and associated P value statistics, see Supplementary Tables 1-3). Stage 2: Enrichment analysis using Pathway Studio to determine overrepresented GO biological processes in the DE miRNA, protein and mRNA lists (see Supplementary Tables 4-6). Stage 3: Integration of three parallel datasets to generate 3 groups of interesting candidates; (1) Potential targets of miRNA translational repression (protein DE without mRNA change and targeted by anti-correlated miRNA): see Table 1 (Downregulated Proteins targeted by Upregulated miRNAs) and Table 2 (Upregulated Proteins targeted by Downregulated miRNAs); (2) Non- microRNA-mediated gene-protein expression (matching mRNA transcripts and proteins were DE, no corresponding anti-correlated microRNA data or results were below detection level): see Supplementary Table 7 and (3) Targets where control of differential protein expression was unclear (protein DE without mRNA or corresponding targeting microRNA change). Stage 4: TargetScan analysis of both groups against miRNAs DE in the opposite direction. Stage 5: Overlap analysis of Supplementary Tables 1-3 with published profiling studies on same cell lines: see Table 3.