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Fig. 6 | Journal of Hematology & Oncology

Fig. 6

From: LincK contributes to breast tumorigenesis by promoting proliferation and epithelial-to-mesenchymal transition

Fig. 6

LincK was physically associated with miR-200 s. a Schematic outlining the predicted binding sites of miR-200 s on LincK. b Dual luciferase reporter assay in 293 T cells co-transfected with microRNAs (NC or miR-200b mimics) and psiCHECK2 (containing wild-type or mutant transcripts). Data were presented as the relative ratio of Renilla luciferase activity to Firefly luciferase activity. c Schematic outline of MS2-Flag RIP assay. 293 T cells were co-transfected by MS2bp-FLAG and MS2bs-LincK or MS2bs. Cell lysates were immunoprecipitated by FLAG M2 beads, and miR-200b endogenously associated with MS2bs-LincK were measured by qRT-PCR. Data were shown as means ± S.D. (n = 3). d RNA pull down assay of the binding of LincK and mir-200b. MCF-7 cell lysates were incubated with biotin-labeled LincK or LacZ, and the mir-200b expression level was detected by qRT-PCR after pull down. RNAs associated with biotin-LincK were compared to that of biotin-LacZ, and U6 was used as non-specific control. Data were shown as means ± S.D. (n = 3). e RIP assay of AGO2 enriched LincK in MCF-7 cells. IgG was used as negative controls. All relative abundances were compared to 1% input. Data were shown as means ± S.D. (n = 3). *p < 0.05; **p < 0.01; and ***p < 0.001 by two-tailed Student’s t test

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