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. 2016 Sep 15;6:33345. doi: 10.1038/srep33345

Figure 4. The binding of PABPs to miRNA intronic AAAAA.

Figure 4

(a) A schematic depiction of forward-mir-517a (AAAAA(+)) or (AAAAA(−)) constructs. (b) A comparison between the expression level of forward-mir-517a (AAAAA(+)) and (AAAAA(−)) in mouse placenta (n = 5–15) and 293T cells (n = 3), determined by qPCR. (c) The expression of PABP family members in mouse or human tissues and cell lines, assessed using standard PCR. Mouse blastocysts were obtained at E3.5, embryos were obtained at E10.5, and placentas at E18.5. Human tissue included term placenta and adult testis. (d) 293T stable cell lines expressed mCherry (control) or mCherry-tagged PABPC1, PABPC4, PABPN1, or SRSF1. TRITC (red), FITC (green), and DAPI (blue) filter sets were used to detect mCherry, EGFP, and DAPI fluorophore, respectively. (e) RIP assays to determine the binding of relevant PABP proteins to the synthesized miR517-RNA ((AAAAA(+) or AAAAA(−)) construct, performed using RT-qPCR as described in Methods (n = 3 for each). *Denotes p < 0.05, **denotes p < 0.01.