Fig. 4

Analysis of the heterologous SUSD2 overexpression in MCF-7.A: Principal component analysis of LC-MS/MS data from the three samples without SUSD2-insert (Ctrl) and carrying the SUSD2 gene in the vector (OE, overexpression). Clone OE2 was analyzed.B: Visualization of a Pearson-correlation based supervised hierarchical clustering. The color legend displays the log2 foldchanges. Analyzed peptides: 24 (PDCD4), 5 (RCN1), 144 (GRP78, HSPA5). For all proteins, the differential expression was > 2 and significant (p < 0.05, Student’s t-test).C: Western Blot analysis of the control cell line and two SUSD2 overexpressing clones for the denoted proteins.D: Quantitative Western Blot analyses of the X-ray films shown in (C). The signals were normalized to α-tubulin, in case of p70 S6 kinase (Thr389) the signals were normalized to total p70 S6 kinase. Detailed information is shown in Supplementary Table S4.E: Response of the cells to treatment with 17β-Estradiol. Cells were cultured in hormone depleted medium (HD) or in presence of 100 nmol/ml 17β-Estradiol (E2). The diagrams show the quantitative analysis of the BCL2 levels (left: control, right: clone OE2). PR A/B: progesterone receptor A/B). D,E: Average values are shown as numbers, and the standard deviation as vertical error bars. The p-values were calculated using Student’s two-sided t-test where p < 0.05 was considered significant. Three biological replicates were analyzed (A-E). Uncropped X-ray film images are shown in supplementary file F1.