Fig. 1: Characterization of hsa_circ_0043280 in CCa cells.

A The top 35 differentially expressed circRNAs in five pairs of CCa tissues and matched adjacent normal tissues GSE102686, were clustered in a Heatmap matrix. B The genomic loci of the hsa_circ_0043280 gene. Hsa_circ_0043280 is synthesized at the TADA2A gene locus containing exons 5 to 8. The back-splice junction of hsa_circ_0043280 was identified by Sanger sequencing. C PCR analysis for hsa_circ_0043280 and its linear isoform TADA2A in cDNA and genomic DNA (gDNA). D qRT-PCR analysis for the expression of hsa_circ_0043280 and TADA2A mRNA after treatment with RNase R in HeLa cells. E qRT-PCR analysis of hsa_circ_0043280 using random primers and oligo dT primers, respectively, in reverse transcription experiments. Hsa_circ_0043280 was notably absent in polyA-enriched samples. F qRT-PCR analysis for the expression of hsa_circ_0043280 and TADA2A mRNAs after treatment with actinomycin D at the indicated time points in HeLa cells. G Cytoplasmic and nuclear mRNA fractionation experiments showing that hsa_circ_0043280 is localized mainly in the cytoplasm of HeLa cells. GAPDH, Cdr1as, and U6 were applied as positive controls in the cytoplasm and nucleus, respectively. H RNA fluorescence in situ hybridization for hsa_circ_0043280 in HeLa cells; the junction probe was complementary to the back-splice junction sequence of hsa_circ_0043280. Nuclei were stained with DAPI. Scale bar, 20 µm. Each experiment was performed at least three times independently. ***P < 0.001.