Figure 6.

circ-FoxO3 interacts with mTOR and inhibits mTORC1 activity

(A) The biotin-labeled circ-FoxO3 probe was incubated in lysates of BMECs with circ-FoxO3 overexpression, and mTOR was identified using an immunoblotting assay. (B) circ-FoxO3 was immunoprecipitated with anti-mTOR-coated magnetic beads, and then detected by qRT-PCR. n = 4. (C) Representative images showed co-localization of circ-FoxO3 and mTOR in BMECs (bEnd.3 and HBMEC). Scale bar, 10 μm. n = 6. White arrowheads, co-localization of circ-FoxO3 and mTOR. (D) The activity of the mTORC1 complex was determined by immunoblotting assay to measure the phosphorylation of S6k1. n = 4. p values indicate one-way ANOVA with Dunnett's multiple comparisons test. OGD/R, oxygen-glucose deprivation for 3 h followed by reoxygenation for 3 h. Data are provided as the mean ± SEM. ∗p < 0.05, ∗∗p < 0.01.