Fig. 8.

MRCCAT1 suppresses NPR3 expression by recruiting Polycomb Repressive Complex 2 to NPR3 promoter. a qRT-PCR analysis of MRCCAT1 in subcellular fraction of Caki-1 cells. U6 and β-actin acted as nucleus and cytoplasm marker, respectively. b RIP assay analysis of the enrichment of MRCCAT1 to EZH2 in 786-O and Caki-1 cells. c Biotinylated MRCCAT1 or antisense RNA were incubated with nuclear extracts of 786-O and Caki-1 cells, targeted with streptavidin beads, and washed. Then the associated proteins were resolved in a gel. Western blot analysis of the specific association of EZH2 and MRCCAT1. d ChIP assays were conducted on NPR3 promoter regions using the indicated antibodies. Enrichment was determined relative to input controls. The results are presented as the mean ± SD from three independent experiments; **P < 0.01, ***P < 0.001 by Student’s t-test