Involvement of the NOTCH1 pathway in the circHECW2-MIR30D-ATG5-mediated EndoMT. (A) LPS induced an increased cleavage of NOTCH1/NICD from NOTCH1. Cells were treated with LPS (10 ng/ml) for different time points. All data are presented as the mean ± SD of 3 independent experiments. *P<0.05, **P<0.01, ***P<0.001 vs. the vehicle control via one-way ANOVA. (B) Transduction of HBMECs with ATG5 siRNA significantly inhibited the increases in the NOTCH1 level induced by anti-MIR30D lentivirus. HBMECs were cotransduced with anti-MIR30D and ATG5 siRNA for 24 h. All data are presented as the mean ± SD of 3 independent experiments. ***P<0.001 vs. the anti-MIRCon cotransduced with the control siRNA group. ##P<0.01 vs. the anti-MIR30D cotransduced with the ATG5 siRNA via one-way ANOVA followed by the Holm-Sidak test. (C) Transduction of HBMECs with circHECW2 siRNA lentivirus significantly inhibited the level of NOTCH1 induced by LPS. HBMECs were transduced with the circHECW2 siRNA or circCon siRNA for 24 h and were subsequently incubated with LPS (10 ng/ml). All data are presented as the mean ± SD of 3 independent experiments. *P<0.05, **P<0.01 vs. the circCon siRNA group via one-way ANOVA. #P<0.05 vs. the LPS-treated circCon siRNA group via one-way ANOVA followed by the Holm-Sidak test. (D) Transduction of HBMECs with the anti-MIR30D lentivirus significantly inhibited the decreases in the NOTCH1 level induced by circHECW2 siRNA. HBMECs were cotransduced with anti-MIR30D and circHECW2 siRNA. All data are presented as the mean ± SD of 3 independent experiments. **P<0.01 vs. the circCon siRNA with anti-MIRCon-cotransduced group. #P<0.05 vs. the circHECW2 siRNA with anti-MIRCon-cotransduced group via one-way ANOVA followed by the Holm-Sidak test. (E) circHECW2, identified as ceRNA, functioned as an endogenous MIR30D sponge to sequester MIR30D and inhibited its activity, which resulted in increased ATG5 expression and consequent EndoMT via the NOTCH1 signaling pathway.