Supplementary Methods and Tables and Figures from DNA Methylation of the SLC16A3 Promoter Regulates Expression of the Human Lactate Transporter MCT4 in Renal Cancer with Consequences for Clinical Outcome

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PDF file, 405K, Supplementary methods, including detailed description of patient cohort, MCT4 immunohistochemical staining and TMA analysis, MCT4 knockdown experiments, Western Blot analysis, RNA isolation and quantification, DNA isolation and promoter methylation analyses, promoter reporter constructs and in vitro methylation, transient transfection and reporter assays as well as statistical analyses. Supplementary Table S1: Comparison of the three independent patient cohorts. Supplementary Table S2: Correlation of MCT4 protein expression in ccRCC tissue with clinicopathological parameters in the TuCP1-cohort. Supplementary Table S3: CpG sites analyzed by the Infinium HumanMethylation450 BeadChip and by MALDI-TOF MS. Supplementary Table S4: Correlation of MCT4 protein expression in ccRCC tissue with clinicopathological parameters in the TuCP2-cohort. Supplementary Table S5: Univariate Cox regression analysis of prognostic factors for survival in the TuCP1-, TuCP2- and the TCGA-cohort. Supplementary Table S6: Results from ROC analysis for survival in the TuCP2- and TCGA-cohort. Supplementary Figure S1: Validation of MCT4 antibody by MCT4 knockdown and representative pictures of stained non-tumor and ccRCC tissue for the evaluation of MCT4 protein expression by the composite IHC score. Supplementary Figure S2: MCT4 protein and mRNA expression and correlation with specific CpG sites in the TuCP2-cohort. Supplementary Figure S3: Kaplan-Meier curve of cancer-specific survival based on percentage of MCT4 protein expression in ccRCC tissue of the TuCP2-cohort. Supplementary Figure S4: DNA methylation at the SLC16A3 promoter in the TCGA-cohort (Infinium Human Methylation 450 Bead Chip) depending on VHL mutations.

Ce résumé expose les affirmations des auteurs. BNTIC ne l’interprète pas comme une validation indépendante des résultats.