Fig. 3

Identification of circHIF1A and its impact on patients’ prognosis. A-C. Identification of circHIF1A. A. qRT-PCR using divergent or convergent primers to detect circHIF1A in LIM1215-R. B. qRT-PCR analysis of circHIF1A and linear mRNA levels in LIM1215-R with or without RNase R treatment. C. Sanger sequencing showing the circular junction sequence of circHIF1A, which is formed by the splicing of the 5’ end of exon 3 and the 3’ end of exon 4. The blue box on the left represents the last 10 nucleotides of the 3’ end of exon 4, and the red box on the right represents the first 10 nucleotides of the 5’ end of exon 3. D. FISH detection of circHIF1A distribution in CRC cells. E-F. FISH detection of the positive (E) and negative (F) expression of circHIF1A in CRC tumor tissues (×400 magnification): the red signals represent circHIF1A labeled with Cy3 probe, and the blue signals represent the cell nuclei labeled with DAPI probe. G. The ORR and DCR of circHIF1A-positive patients and negative patients. H. The mPFS of circHIF1A-positive patients and negative patients. ^*P < 0.05, ^**P < 0.01, ^***P < 0.001