circLPAR3 is verified and characterized in docetaxel-resistant prostate cancer cells. (IMAGE)
Caption
(A) Venn plot of the differentially expressed docetaxel-resistant circRNAs. (B) The expression of circLPAR3 was detected by quantitative reverse transcription PCR in docetaxel-resistant prostate cancer cells (22Rv1-DR, DU145-DR, and PC3-DR) and sensitive cell lines (22Rv1, DU145, and PC3). (C) Schematic illustration of circLPAR3 formation via the circularization from exon 2 in the LPAR3 gene. (D) The expression of circLPAR3 in cDNA and gDNA of 22Rv1 and DU145 cells were confirmed by quantitative reverse transcription PCR with the divergent and convergent primers. (E) The relative expression of circLPAR3 and LPAR3 was measured by quantitative reverse transcription PCR after treatment with RNase R. (F) Cells were treated with actinomycin D (2 μg/mL) and harvested after 0, 4, 8, 12 and 24 h. Relative expression of circLPAR3 and LPAR3 was calculated and normalized to 0 h to compare the RNA stability. (G) Nuclear–cytoplasmic fractionation assay was used to assess the intracellular location of circLPAR3 in drug-resistant and parent cells. U6 was considered as a nuclear control and β-actin was used as a cytoplasmic protein control. (H) The subcellular location of circLPAR3 was also evaluated by fluorescence in situ hybridization assays. Nuclei were stained with DAPI. ∗∗P < 0.01, ∗∗∗P < 0.001, and ∗∗∗∗P < 0.0001; ns, nonsignificant.
Credit
Yetao Zhang, Yuxiang Dong, Kai Li, Tong Zhao, Yongshan Li, Mingyang Pang, Yong Wei, Bing Yao, Qingyi Zhu
Usage Restrictions
Credit must be given to the creator. Only noncommercial uses of the work are permitted. No derivatives or adaptations of the work are permitted.
License
CC BY-NC-ND