Fig. 1

Identification and characterization of circ_001422 in OS. a. Cluster heatmap showing the top ten most upregulated and downregulated circRNAs identified by high-throughput sequencing in 3 paired OS tissues and adjacent noncancerous tissues. The rows show circRNAs, while the columns show samples. The red and green strips indicate upregulated and downregulated circRNAs, respectively. The list of the circRNAs is presented as Chr:5′ nt position-3′ nt position and the type of the strand (positive or negative). b. Schematic illustration of circ_001422 formation and the results of Sanger sequencing. c. Relative quantification (RQ) value of circ_001422 in OS cells and normal osteoblasts. d. RQ values of circ_001422 and NSD2 mRNA in OS cells under RNase R treatment. e. The linear and back-splicing products were amplified with convergent and divergent primers, treated with and without RNase R, and subjected to PCR. f. RQ values of circ_001422 and NSD2 mRNA after treatment with actinomycin D at the indicated time points. g & h. Nucleocytoplasmic fractionation and FISH revealed the subcellular localization of circ_001422 in OS cells. Nuclei were stained with DAPI, and circ_001422 probes were labeled with Cy3. Scale bar, 50 μm. n.s., not significant; ^*P < 0.05; ^**P < 0.01; ^***P < 0.001