Fig. 5

PEBP1P2 mediates KLF13 by post-transcriptional regulation. a, b Following transfection with indicated shRNAs or overexpressing vectors, the RNA levels of miR-296,-616, and 3194 were assessed by real-time PCR, respectively. c, d By real-time PCR, RNA levels of PEBP1P2 were determined after transfection with mimics or inhibitors. e, f Following transfection with the indicated mimics or inhibitors, we detected the levels of miR-296/-616/-3194 by real-time PCR. g Assay model for AGO2-RIP. h AGO2 antibody was used in the RIP assay, then the enrichment of PEBP1P2 was detected with real-time PCR. i Assay model for MS2-RIP. j RNA derived from MS2-RIP was examined by real-time PCR, with the enrichment fold-changes normalized to IgG control RNA. k The miRNAs-Biotin complex was enriched by Biotin-antibody with protein extracts, and the enrichment of PEBP1P2 was measured by real-time PCR. l-m Transfection of HEK 293 T cells with luciferase reporter vectors and miRNA mimics was performed, and luciferase reporter activity was detected. n Diagram showing the selection of direct downstream targets of miR-296/-616/-3194. o-p By real-time PCR, the levels of mRNA of KLF13 were determined after transfection with indicated mimics, inhibitors, and lentiviruses. q AGO2 antibody was used to perform RIP assays, and real-time PCR was then used to detect KLF13 mRNA enrichment. r RNA derived from MS2-RIP was examined by real-time PCR, with fold changes of enrichment normalized relative to IgG control. s By using Biotin-antibodies in combination with protein extracts, miRNAs and biotin complexes were enriched, and the enrichment of KLF13 mRNA was determined by real-time PCR. The data are presented as the mean ± SD, ***P < 0.001