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. 2020 Mar 10;11:212. doi: 10.3389/fgene.2020.00212

FIGURE 4.

FIGURE 4

Hsa_circ_0097435 acted as a sponge for multiple miRNAs. (A) The potential binding sites of miRNAs related to hsa_circ_0097435 are shown. (B) Detection of transient, efficient expression of C97435-MS2 (green fluorescence), Scale bars, 100 μm. (C) Detection of efficient expression of MS2-CP (red fluorescence), Scale bars, 100 μm. (D) Compared with the negative-control group, 5 miRNAs in the circRNA-enrichment group were upregulated, including hsa_miR_6799_5P (**p < 0.01), hsa_miR_5000_5P (**p < 0.01), hsa_miR_609 (**p < 0.01), hsa_miR_1294 (**p < 0.01), and hsa_miR_96_5P (**p < 0.01). (E) WB analysis of pulldown products. The AGO2 protein was significantly pulled down by hsa_circ_0097435. The amount of AGO2 protein in the C97435-MS2 pulldown group was significantly higher than in the C97435 pulldown group. (F) Histogram showing quantitative comparison of the polypeptide TTPQTLSNLCLK between the sample groups. AGO2 immunoprecipitation was followed by mass spectrometry, and 3–8 fragment ions of the specific peptide TTPQTLSNLCLK were selected for quantitative analysis. (G) Immunoprecipitation of AGO2 from AC16 cells. Compared with the IgG group, 4 miRNAs in the AGO2 group were upregulated, including hsa_miR_6799_5P (**p < 0.01), hsa_miR_5000_5P (**p < 0.01), hsa_miR_609 (**p < 0.01), and hsa_miR_1294 (**p < 0.01).