Fig. 5

Molecular profiling and functional analysis reveal a role of ADAM10 in cell cycle progression and apoptosis. A-E Proteome and secretome analysis of ADAM10 KO and control SEM and Nalm-6 cells. A Workflow of mass spectrometry-based secretome and proteome analysis of ADAM10 KO and CTRL cells. B SEM proteome. Heat map of unsupervised hierarchical clustering of significantly regulated proteins of control (n = 4) vs. ADAM10 KO (n = 8) in SEM cells (two-sample test, permutation-based FDR < 0.05). C Pathway enrichment results of the proteome analysis described in B. The five most significantly altered pathways are depicted. FDR: false discovery rate. D SEM secretome. Heat map of unsupervised hierarchical clustering of significantly regulated secreted proteome of control (n = 4) vs. ADAM10 KO (n = 4) in SEM cells (two-sample test, permutation-based FDR < 0.05). E Box plots showing significantly regulated secreted proteins in SEM cells with ADAM10 KO. Plot displays z-scored log 2 protein intensity of selected proteins. F Transcriptome analysis of ADAM10 KO (n = 3) and CTRL PDX (n = 4) samples of ALL-199 and ALL-265. Heatmap of genes differentially expressed between ADAM10 KO and CTRL cells with unadjusted p value of ≤ 0.05 and fold change < 0.5 or > 2 is shown. G Pathway enrichment results of transcriptome analyses described in F were mapped into a network of gene sets (nodes) related by gene overlap (lines). Node size is proportional to the number of genes in each set and the enrichment significance (FDR p value) is represented as a node color gradient. Proportion of shared genes between gene sets is depicted as the thickness of the blue line surrounding the nodes. The major functional groups are annotated. Data analyzed and visualized by GSEA 4.1.0 and Cytoscape 3.9.0. H Gene set enrichment analysis (GSEA) for the KEGG term cell cycle (p < 0.005 and FDR q value < 0.33, Norm p = 0.01). I Proteome of PDX cells. Heatmap of significantly regulated proteins by unsupervised hierarchical clustering of ALL PDX sample with control sgRNA (ALL-199 n = 4, ALL-265 n = 4) vs. ADAM10 KO sgRNA (ALL-199 n = 5, ALL-265 n = 4) (two-sample test, permutation-based FDR < 0.05). J Plot displaying the enriched and de-enriched GO term categories (for panel I) upon ADAM10 KO in ALL PDX samples by Fisher’s exact test. K Cell cycle analysis of ADAM10 inhibitor (GI254023X, 490 µM)- or DMSO-treated ALL-199 PDX cells. Percentage of cells in the indicated cell cycle phase was quantified on day 1, day 2 and day 3 of treatment with the ADAM10 inhibitor or DMSO. Each dot represents the mean of four replicates. G1 = Gap phase 1, S = Synthesis phase, G2/M = Gap phase 2/mitosis. ** p < 0.01, * p < 0.05 by paired t-test. L Apoptosis assay in ALL-199 PDX cells with ADAM10 KO (n = 3) or treated with ADAM10 inhibitor (GI254023X, 490 µM, n = 6). *** p < 0.001, * p < 0.05 by paired t-test