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Fig. 4 | Journal of Animal Science and Biotechnology

Fig. 4

From: NORHA, a novel follicular atresia-related lncRNA, promotes porcine granulosa cell apoptosis via the miR-183-96-182 cluster and FoxO1 axis

Fig. 4

NORHA is a molecular sponge of the miR-183-96-182 cluster. a An interaction network between NORHA and miRNAs that were downregulated in EAF. The data of miRNA-Seq are not shown. The red triangle represents NORHA, which was upregulated in EAF, and blue ellipses represent the miRNAs that were downregulated in EAF. b Chromosomal localization of miR-183, miR-96, and miR-182 show that they form a miRNA cluster in a common genome region. c Schematic showing the interactions of miR-183, miR-96, and miR-182 seed sequences with the NORHA RNA sequence. d Minimum free energy (MFE) of miR-183, miR-96, and miR-182 binding to NORHA was predicted by RNAhybrid. e Schematic showing the reporter constructs of NORHA. The NORHA sequence containing wild-type and mutant miRNA-binding sites was amplified and inserted into a prim-GLO plasmid to construct a luciferase reporter vector. f, g Wild-type (f) or mutant (g) reporter vectors were cotransfected with miR-183 mimics (mi), miR-96 mimics (mi), and miR-182 mimics (mi) into HEK293T cells, and luciferase activity was measured. h RIP assay was performed with an anti- AGO2-specific antibody. The levels of NORHA enriched on the AGO2 protein were detected by qRT-PCR. IgG antibody was used as a negative control. i The levels of miR-183, miR-96 and miR-182 in GCs with NORHA knocked down. The data are presented as the mean ± SEM of at last three independent experiments. * P < 0.05. ** P < 0.01. ns, not significant

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