Figure 5.

Hsa_circ_100791 acts as a sponge for miR-487b-5p to regulate TRIM13 in HNEpCs. (A) The databases predicted the sequences of hsa_circ_100791, which are complementary to miR-487b-5p seed sequence. (B and C) The luciferase reporter assays results showed that the luciferase activity of wild-type version significantly reduced when miR-487b-5p mimics in HNEpCs, while enhanced when miR-487b-5p knockdown in HNEpCs. But, the luciferase activity of mutant version was obviously different. (D and E) FISH assay revealed that hsa_circ_100791 (red) localized with miR-487b-5p (green) in the cytoplasm. Nuclei were stained with DAPI. Hsa_circ_100791 and miR-487b-5p were labeled with Cy3 and FAM, respectively. (F) qRT-PCR analysis result showed that the expression mRNA level of TRIM13 was significantly enhanced in hsa_circ_100791 overexpression group, but obviously reduced in hsa_circ_100791 knockdown group. (G and H) Western blot analysis result showed that the expression protein level of Trim13, p65 and p-p65 was significantly increased in hsa_circ_100791 overexpression group, whereas decreased in hsa_circ_100791 knockdown group. Data were presented as mean ± SEM. N=3 for each group.

Abbreviations: FISH, Fluorescence in situ hybridization; SEM, standard error of mean.