Fig. 4

SOX9 is the downstream gene mediated by circEHD2. A, The cluster heat maps showed the differentially expressed genes regulated by circEHD2 based on next-generation sequencing (NGS) analysis. The red and blue strips indicate up-regulated and down-regulated genes, respectively. B and C, qRT–PCR analysis of the cAMP signal pathway-related genes in OSRC-2 (B) and 786-O (C) cells with circEHD2 knockdown. D and E, qRT–PCR analysis of the expression level of SOX9 when downregulation (D) or upregulation (E) of circEHD2 in OSRC-2 and 786-O cells. F and G, Western blot showed SOX9 expression level in whole protein extracts (F) and nuclear protein extracts (G) of OSRC-2 cells when silencing circEHD2. H, Representative images of IHC and ISH showed the expression of SOX9 and circEHD2 in ccRCC tissues and paired NATs (n = 80). Scale bars: 50 μm. I, The H-score of SOX9 in ccRCC tissues and paired NATs (n = 80). J, The Pearson correlation analysis showed that circEHD2 expression level was positively correlated with SOX9. Error bars, standard deviation (SD) of three independent experiments. *, P < 0.05; **, P < 0.01; ***, P < 0.001