Fig. 5

ROR abolishes histone H3K9 methylation of the TESC promoter. a The location of mature ROR. ROR was mainly present in the nucleus. U2 RNA was used as a positive control for nuclear RNA. b Schematic diagram of the TESC promoter region and ROR lncRNA. T7 through T12 and S1 through S6: primer names, arrow: transcriptional direction, pTESC-1 and pTESC-2: two different biotinylated TESC promoter fragments; pTESC-3: biotinylated DNA fragments 10 kb upstream of TESC. Sites a, b, and c: different detecting locations of RNA IP. c The interaction of ROR and the TESC promoter. The TESC promoter (pTESC-1 and pTESC-2) specifically interacts with exon 3 of the ROR (site b). pTESC-3 was used as a negative control locus. Fibroblasts and lncRNA-KCNQ1OT1 were used as negative controls. Input: total RNA was reverse transcribed before incubation with labeled pTESC fragments and amplified with GAPDH primers. d TaqMan real time-PCR used to quantify the enrichment of ROR on the TESC promoter. TESC binds near exon 3 of ROR (site b). pTESC-3 was used as a negative control locus. KCNQ1OT1 was used a negative control. e-g ChIP assay detecting H3K9 trimethylation of the TESC promoter. Sites X, Y, and Z: different sites used in this assay. IgG: native control. All data are presented as the means ± SD. *P <0.05: compared with the control and mock