Supplementary Data from Serum- and Glucocorticoid-induced Kinase Sgk1 Directly Promotes the Differentiation of Colorectal Cancer Cells and Restrains Metastasis

Supplementary Table 1. Differentially expressed genes from colonic crypt tops vs. bottoms of normal individuals (adjusted p<0.05). Supplementary Table 2. SGK1 expression of CRC cell lines from Broad Institute, The Cancer Cell Line Encyclopedia. Supplementary Table 3. Summary patient characteristics for sporadic CRC microarray. Supplementary Figure 1. Expression of SGK1 mRNA in normal colon (n=40), CRC (n=217) and CRC lines (n=7). Supplementary Figure 2. Average SGK1 expression levels resulting from the Tet-regulated, lentivirus-based system (qRT-PCR dCt values normalised to a GAPDH control). Supplementary Figure 3. SGK1 protein levels in representative anti-FLAG western blots of HT29 and LS174T cells transduced with FLAG-tagged SGK1-TetOff virus. Supplementary Figure 4. Increased levels of SGK1 induce degradation of the target protein Nedd4-2, confirming functionality of the transduced SGK1 protein. Supplementary Figure 5. Microscopic effects of re-expression of SGK1 in HT29 cells. Panels (a) to (d) show division of wild type cells (yellow arrows). Panels (e) to (h) show division of HT29 SGK1 re-expressing cells. Supplementary Figure 6. Representative images of immunofluorescence analysis showing ZO-1 and β-catenin expression in SGK1 re-expressing LS174T cells and controls. Supplementary Figure 7. Representative western blots demonstrating reduced MYC protein expression following SGK1 re-expression in SW480 (p=0.04, t test) Supplementary Figure 8. PKP3 induced by SGK1. Supplementary Figure 9. Effects of SGK1 on primary tumour growth and lung metastasis in an orthotopic transplant model Supplementary Figure 10. Representative immunohistochemistry section demonstrating SGK1 expression using EMD Millipore anti-SGK1 07-315. Supplementary Figure 11. Overall survival of CRC patients by SGK1 expression. Data are from The Cancer Genome Atlas, mRNAseq data (RSEM normalised)