Research Paper Volume 12, Issue 8 pp 7232—7247

Figure 2. The interaction between XIST and miR-101a-3p. (A) Bioinformatic analysis (StarBase and miRbase) indicated that miR-101a-3p was a target of XIST (left panel) and the luciferase activity was analysed (right panel) (n=3). XIST-WT: wild-type XIST, XIST-Mut: XIST with mutated 3’-UTR. (B) In vitro transfection efficiency was determined by measuring miR-101a-3p and miR-101a-3p inhibitor using qRT-PCR (n=3). (C) The results of RNA pull-down assay. NMCMs cell lysates were incubated with biotin-labeled miR-101a-3p, after pull-down, microRNAs was extracted and analysed by qRT-PCR (n=3). (D) The results of RNA immunoprecipitation assay. Anti-AGO2 RIP was performed in NMCMs cells transiently overexpressing miR-101a-3p mimic or nc mimic, followed by qRT-PCR to detect XIST associated with AGO2 (n=3). nc mimic: a sequence did not interact with XIST; con: control. Data are shown as mean ± SD, * P<0.05.