Fig. 3

TGFBR2 is the GABPA target gene. A GABPA and TGFBR2 expression is highly correlated with each other in the TCGA ccRCC tumors. B The GSEA analysis reveals that the TGFβ signaling is enriched in ccRCC tumors expressing higher GABPA in the TCGA cohort of ccRCC. C The volcano plot shows TGFBR2 among the top downregulated genes in GABPA-depleted cells, as determined using RNA seq derived from 3 independent experiments (Exp). D The heatmap of RNA seq results illustrates significantly downregulated expression of the TGFβ pathway factors. E The GSEA analysis of RNA seq results shows the diminished TGFβ pathway enrichment in GABPA-depleted cells. F Venn diagram reveals five overlapped genes based on the integrated analyses of TCGA, RNA seq and GABPA ChIP-seq of leukemic and liver cancer cells. G GABPA knockdown downregulates while its overexpression upregulates TGFBR2 expression. H Immunofluorescence results further reveal diminished TGFBR2 expression in GABPA-depleted cells. Scale bars: 10 µm. I Top panel: The schematic drawing shows the GABPA binding motifs on the TGFBR2 promoter and mutated nucleotides. Bottom panel: GABPA knockdown inhibits while its overexpression stimulates the TGFBR2 promoter activity. J Top panel: The schematic diagram shows the primer locations spanning GABPA sites for ChIP assay. Bottom panel: The enrichment of GABPA on the TGFBR2 promoter, as determined using ChIP assay. Three independent experiments were performed. *, ** and *** denote P < 0.05, 0.01 and 0.001, respectively