Fig. 4. SDNOR, a novel antiapoptotic lncRNA, is regulated by SMAD4.
Detection of 5′ and 3′ terminals of the novel lncRNA by using 5′RACE (a) and 3′RACE (b). c Schematic annotation of SDNOR with associated UCSC Genome Browser tracks depicting genomic locus, GC percent, H3K27Ac modification as well as mammalian conservation. d qRT-PCR validation of SDNOR expression levels in pig ovarian GCs and TCs (Theca cells). e Detection of SDNOR expression in healthy follicles (HF) and atresic follicles (AF), assessed by qRT-PCR (n = 10). f SDNOR expression levels in porcine GCs after SDNOR knockdown, determined by qRT-PCR. g FACS analysis (left) and quantification (right) of the apoptosis rate of porcine GCs after SDNOR knockdown. h Representative western blot analysis (left) and quantification (right) of c-Caspase-3 protein levels in GCs after SDNOR knockdown. i Luciferase activities of reporter vectors containing pig SDNOR promoter in porcine GCs with or without SMAD4 overexpression (S4OE) were assessed. j Diagram depicting SDNOR promoter (top) and ChIP assay (down), SBE shown as pink diamonds; F1′/R1′, F2′/R2′, and F3′/R3′: primers used for ChIP and ChIP-qPCR assay. k ChIP-qPCR were performed to detect the different SBEs in the promoter of SDNOR following immunoprecipitation with a SMAD4 antibody. Throughout, all experiments were performed in triplicate. Data in d–k are shown as mean ± S.E.M. (n = 3, each). **p < 0.01 versus control or scrambled.