Fig. 4: NOLC1 mediates the regulation of Runx2 in ccRCC cell growth and metastasis.

A RNA-sequencing was performed on ACHN cells transfected with two siRNA targeting Runx2, and the genes with significant changes were indicated by heat map. B The mRNA expressions of Runx2 and NOLC1 in ACHN and 786-O cells after knockdown of Runx2 were analyzed by qRT-PCR. C Western blot analysis showed the downregulation of NOLC1 in SKRC39 and CAKI-1 cells after overexpression of Runx2. β-Actin was also tested as a loading control. D Luciferase report assay showed that high expression of Runx2 inhibited the transcription of NOLC1. E IHC staining analysis in ccRCC tissue array showed the negative correlation between the expressions of Runx2 and NOLC1. F qRT-PCR was used to analyze the expression level of NOLC1 in ccRCC and corresponding normal renal tissues at mRNA level. G Survival analyses of ccRCC patients with different expression level of NOLC1 based on TCGA datasheet. H–J MTT test (H), EdU incorporation (I), and foci formation (J) assays were performed to analyze the proliferation of 786-O and ACHN cells after knockdown of NOLC1 or/and Runx2, respectively. K The migration ability of ccRCC cells after interference of NOLC1 or/and Runx2 was analyzed by Transwell migration assay. In all panels, *, P < 0.05; **, P < 0.01; ***, P < 0.001.