Figure 7.

circ-FoxO3 sequesters E2F1 to inhibit mTOCR1 activity and promotes autophagy

(A) circ-FoxO3 was immunoprecipitated with anti-E2F1-coated magnetic beads, and then detected using qRT-PCR. n = 3. p values indicate one-way ANOVA with Dunnett's multiple comparisons test. (B) The localization of circ-FoxO3 and E2F1 in BMECs (bEnd.3 and HBMEC) was determined by FISH and IF. n = 5. Scale bar, 10 μm. The dotted line is the boundary of the nucleus. White arrowheads show the co-localization of circ-FoxO3 and E2F1 in cytosol. Yellow arrowheads indicate the E2F1 in the nucleus. (C) Quantitative integrated optical density of E2F1 in nucleus in (B). p values indicate a two-tailed unpaired Student's t test. (D) The levels of E2F1, PFKFB3, p-S6K1, S6K1, SQSTM1, and LC3B were measured by western blotting. n = 3. (E) Quantitative results of the bands in (D). n = 3. p values indicate one-way ANOVA with Dunnett's multiple comparisons test. (F) The schematic diagram of circ-FoxO3 in the attenuation of BBB damage during I/R. OGD/R, oxygen-glucose deprivation for 3 h followed by reoxygenation for 3 h. I/R, ischemia/reperfusion. Data are provided as mean ± SEM. ∗p < 0.05, ∗∗p < 0.01.