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Fig. 2 | Molecular Cancer

Fig. 2

From: circPTEN1, a circular RNA generated from PTEN, suppresses cancer progression through inhibition of TGF-β/Smad signaling

Fig. 2

The identification and characteristics of circPTEN1 in colon cancer. A. Schematic illustration showing the PTEN exon 1 (partial)-exon 5 circularization forming circPTEN1. The presence of circPTEN1 was validated by RT-PCR, followed by Sanger sequencing. Black arrow represents “head-to-tail” circPTEN1 splicing sites. B. The presence of circPTEN1 was validated in DLD1 and LoVo cell lines by RT-PCR. Divergent primers amplified circPTEN1 in cDNA but not in genomic DNA. β-actin was used as a negative control. C. Northern blotting analysis of circPTEN1 and PTEN mRNA levels in LoVo cells by hybridization with exon 5 (top, left) and exon 5-exon 1 junction (top, right) probes with and without RNase R treatment. GAPDH mRNA with or without RNase R treatment was detected as a control. D. qRT-PCR analysis of the expression of circPTEN1 and PTEN mRNA after treatment with RNase R in DLD1 and LoVo cells. ****, P < 0.0001. E. qRT-PCR for the abundance of circPTEN1, PTEN mRNA and GAPDH mRNA in DLD1 and LoVo cells treated with Actinomycin D at the indicated time points. F. The levels of circPTEN1 in the nuclear and cytoplasmic fractions of DLD1 and LoVo cells. G. FISH detection of circPTEN1 in colon cancer cells. Nuclei were stained with DAPI. Scale bar, 10 μm

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