Fig. 3.

CircFOXO3 may function as a miR-138-5p and miR-432-5p sponge. (A) CircFOXO3 was detected by FISH in U87-MG and U251-MG. Nuclei were stained with 4′,6′-diamidino-2-phenylindole. Scale bar = 20 μm. (B) A schematic drawing showing the putative miR-138-5p/miR-432-5p binding sites with respect to circFOXO3. (C, D) MiR-138-5p/miR-432-5p expression increased upon circFOXO3 KD and decreased after circFOXO3 OE. (E, F) CircFOXO3 expression decreased upon ectopic miR-138-5p/miR-432-5p expression and increased after miR-138-5p/miR-432-5p inhibition. (G) Biotinylated wild-type/mutant miR-138-5p/miR-432-5p were transfected into T98G cells overexpressing circFOXO3. After streptavidin capture, circFOXO3 expression was detected by qRT-PCR. (H) Luciferase reporter assays were reported in HEK293T cotransfected with reporter plasmid (or the corresponding mutant reporter) and the indicated miRNAs. (I) RIP experiments were performed using an anti-AGO2 antibody and U87-MG cells. A multiple comparisons test adjusted P-value of <0.05 was considered statistically significant. Error bars represent the mean ± SD.