Fig. 5.

CRNDE promoted DACT1 expression by increasing H3K27 acetylation in chondrocyte‑like ATDC5 cells. A Using UCSC and ENCODE online databases, H3K27ac was found to be highly enriched in the DACT1 promoter region. B Cytoplasmic & Nuclear RNA Purification Kit was used to isolate cytoplasmic and nuclear RNA. And the expression of CRNDE in the nucleus and cytoplasm was evaluated using qPCR. U6 and GAPDH were used as nuclear and cytoplasmic markers, respectively. C FISH analysis (Bar = 2 cm) of the enrichment level of CRNDE in nucleus of chondrocyte‑like ATDC5 cells. D ChIP assay performed in chondrocyte‑like ATDC5 cells showed the enrichment of H3K27ac in the DACT1 promoter region. E Chondrocyte‑like ATDC5 cells were treated by IL-1β for 24 h, and then the the enrichment of H3K27ac in the DACT1 promoter was analyzed using ChIP assay. F, G After overexpression or interference with CRNDE in chondrocyte‑like ATDC5 cells, the enrichment of H3K27ac in DACT1 promoter was detected using ChIP assay. H, I Chondrocyte‑like ATDC5 cells were infected with Ad-CRNDE for 48 h, and then a histone acetyltransferase inhibitor (C646, 25 μM) was used to treat cells for 1 h. The mRNA and protein levels of DACT1 were determined. The sample size was n = 5, and each test was independently repeated at least three times. Student’s t test was used to detect differences between two groups. *P < 0.05, **P < 0.01