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. 2022 May 3;10:850708. doi: 10.3389/fcell.2022.850708

FIGURE 4.

FIGURE 4

CircFGGY promotes the expression of Smad7 by competitively targeting miR-545-3p. (A,B) qRT-PCR and Western blot detected the RNA and protein expression of Smad7 in MHCC97H cells transfected with circFGGY overexpression vector and HepG2 cells transfected with ASO-circFGGY. (A): data were shown as mean ± SD, n = 3. **p < 0.01, ***p < 0.001 in independent Student’s t test. (C) Venn diagram showing potential miRNA targets for circFGGY and SMAD7. The miRNA targets for circFGGY were obtained from circbank database. The miRNA targets for Smad7 were obtained from targetscan and miRDB database. The HCC associated miRNAs were obtained from MNDR database. (D) The luciferase activity of the wild-type or mutant circFGGY/Smad7 after co-transfection with miR-545-3p mimics or NC-mimics in MHCC97H cells. Data were shown as mean ± SD, n = 3. **p < 0.01, ***p < 0.001 in independent Student’s t test. Wt, wild-type; mt, mutant. (E) In situ hybridization assay showing colocalization of endogenously expressed circFGGY and miR-545-3p in the cytoplasm. The circFGGY probes were labeled with cy3. The miR-545-3p probes were labeled with FITC. Nuclei were stained with DAPI. Scale bar, 20 µm. (F) qRT-PCR detected the RNA expression of miR-545-3p and Smad7 in the 50 paired HCC and adjacent normal tissues. Wilcoxon matched-pairs signed rank test was used. Data represented as median (Min, Max), n = 50. ***: p < 0.001. (G) Pearson correlation analysis between RNA expression of Smad7 and circFGGY/miR-545-3p in clinical tumor tissues. n = 50.