Research Paper

Figure 2. Circ-DNAH1 is derived from the DNAH1 gene exon 67-70 and predominately localized in the cytoplasm. (A) Schematic illustration of the generation of circ-DNAHI from the exons 67 to 70 of the DNAH1 genome by back splicing. The immediate DNA sequence from the splice donor and splice acceptor sites was indicated (left), and was confirmed by the data from RNA sequencing (right). (B, C) qRT-PCR analysis of circ-DNAH1 and DNAH1 mRNA expression in MG63 and U2OS cells treated with Actinomycin D for different time (0 h, 4 h, 8 h, 12 h and 24 h). (D, E) Quantification of circ-DNAH1 and DNAH1 mRNA in total RNA of MG63 and U2OS cells digested with or without RNase R. (F, G) qRT-PCR analysis of the abundance of circ-DNAH1 and DNAH1 mRNA in the nuclear and cytoplasmic fractions isolated from MG63 and U2OS cells, (H) FISH analysis of circ- DNAH1 in the MG63 and U2OS. U6 and 18S RNA were stained by fluorescent probe as well. Data we shown as mean ± SD. **P < 0.01 and ns = no significance.