Fig 1: MSI1 was a downstream target of miR-296-5p in CRC cells. (A) The binding sequence between MSI1 and miR-296-5p was predicted by starBase. (B and C) SW480 and SW620 cells were co-transfected with MSI1-WT or MSI1-MUT and miR-NC or miR-296-5p, and then the luciferase activity was measured through dual-luciferase reporter assay. (D and E) The expression levels of miR-296-5p and MSI1 mRNA was determined by qRT-PCR in SW480 and SW620 cells transfected with miR-NC, miR-296-5p, anti-miR-NC, or anti-miR-296-5p. (F) The protein abundance of MSI1 was detected by Western blot analysis in SW480 and SW620 cells transfected with miR-NC, miR-296-5p, anti-miR-NC, or anti-miR-296-5p. (G and H) The mRNA and protein expression of MSI1 in CRC cells (SW480 and SW620) and FHC cells were analyzed by qRT-PCR and Western blot analyses, respectively. *P<0.05.
Fig 2: MiR-296-5p suppressed tumorigenesis and increased radiosensitivity by targeting MSI1 in CRC cells. SW480 and SW620 cells were transfected with miR-NC, miR-296-5p, miR-296-5p + pcDNA, or miR-296-5p + MSI1. (A and B) MSI1 mRNA and protein expression were measured by qRT-PCR and Western blot analyses, respectively. (C and D) Cell cycle distribution was detected using flow cytometry analysis. (E and F) Cell proliferation was evaluated by MTT assay. (G and H) Colony formation assay was applied to determine cell survival fraction in SW480 and SW620 cells exposed to different doses of radiation. (I and J) Cell apoptosis was examined by flow cytometry analysis in SW480 and SW620 cells irradiated with 0 Gy or 6 Gy. (K and L) Western blot assay was conducted to detect the protein levels of Bcl-2 and Bax in SW480 and SW620 cells irradiated with 0 Gy or 6 Gy. *P<0.05.
Fig 3: Exosomal circ_IFT80 facilitated tumor growth in vivo. SW480 cells were inoculated subcutaneously into the nude mice. 1 week later, Exo-circ_IFT80 or Exo-Vector was subsequently injected into mice. (A) Tumor volume was monitored every week. (B) Tumor weight was measured 4 weeks later. (C–E) The expression levels of circ_IFT80, miR-296-5p and MSI1 mRNA were determined by qRT-PCR in the collected tissues. (F) MSI1 protein abundance in the collected tissues was detected by Western blot assay. *P<0.05.
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