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. 2020 Aug 13;2020:5103272. doi: 10.1155/2020/5103272

Figure 3.

Figure 3

miR-1200 directly bound to circ_0026359. (a, b) Location of circ_0026359 in SGC-7901/CDDP and MKN-45/CDDP cells was examined by nuclear and cytoplasmic separation assay. circ_0026359 was detected by RT-qPCR. U6 and GAPDH were examined as control. (c) Predicted consequential pairing of circ_0026359 with miR-1200 and the mutant consequence of circ_0026359 used in luciferase reporter assay. (d, e) Luciferase reporter assay of SGC-7901/CDDP and MKN-45/CDDP cells cotransfected with miR-1200 mimics/negative control miRNA (miR-NC) and luciferase reporter plasmid pmirGLO containing wild-type circ_0026359 sequence (hsa_circ_0026359-wt)/mutant circ_0026359 sequence (hsa_circ_0026359-mt). (f) RT-qPCR was carried out to examine miR-1200 levels in SGC-7901/CDDP and MKN-45/CDDP cells after transfection with si-circ_0026359#1, si-circ_0026359#2, or siNC. U6 was detected as control. (g, h) RNA pull-down assay: (g) circ_0026359 and (h) miR-1200 were enriched by biotinylated circ_0026359 probe, and their levels were examined by RT-qPCR. ∗P < 0.05; ∗∗P < 0.01. ns: no significance.