Fig. 3

Nuclear NORSF is an endogenous ceRNA of miR-339 in sow GCs. a LOC102164325 is located in a QTL region for sow fertility trait. The information on this QTL is obtained from the pig QTLdb database (https://www.animalgenome.org/cgi-bin/QTLdb/SS/index). b-c miR-339 levels in sow GCs treated with a NORSF-siRNA (b) and an overexpression vector pcDNA3.1-NORSF (c), were determined by qPCR. Data were normalized to U6 levels. n = 3. d NORSF levels in GCs treated with miR-339 mimics were determined by qPCR. Data were normalized to GAPDH mRNA levels. n = 3. e In vivo RNA pull-down assay. GCs were cross-linked, NORSF-RNA complexes were pulled down by using biotinylated probes for NORSF, and miR-339 levels were determined by qPCR. The group without the addition of biotin-labeled probes is used as negative control. PD, pull-down. n = 3. f Schematic showing miR-339 MRE in the pig CYP19A1 PII promoter. The mfe is evaluated by RNAhybrid. g NORSF induces CYP19A1 promoter activity via miR-339. Luciferase assays were performed using a DLR™ assay system. n = 3. h-i NORSF induces CYP19A1 transcription in GCs via miR-339. After pcDNA3.1-NORSF and miR-339 mimics co-transferred into GCs for 48 h, CYP19A1 mRNA (h) and protein (i) levels were detected by qPCR and western blotting, and normalized by GAPDH mRNA and protein levels, respectively. n = 3. Values are means ± SEM. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, no significant