Fig 2: LY294002 (a PI3K inhibitor) inhibited the phosphorylation of PI3K, AKT, and VEGFR2 in follicle microvascular endothelial-like cells of chicken (FMECs) induced by Angelica sinensis (AS) extract. ß-actin was used as an internal reference protein. All experiments were performed in triplicate, and the data are the mean ± S.E.M (*P < 0.05, **P < 0.01, ***P < 0.001).

Fig 3: NGR1 resumed the decreased MTNR1A in the hippocampus induced by SD. (A) Immunostaining of MTNR1A in the hippocampus. Scale bar: 200 and 50 µm. (B) Comparison of immunofluorescence intensity of MTNR1A in the hippocampus. n = 3/group. (C) Protein expression of MTNR1A in hippocampi. n = 5/group. (D–G) Protein expressions of the phosphorylated PI3K, Akt, and mTOR in hippocampi. n = 5/group. *p < 0.05; ***p < 0.001 vs. control group. ##p < 0.01, ###p < 0.001 vs. SD group. (NGR1: notoginsenoside R1; MTNR1A: melatonin receptors 1A; SD: sleep deprivation; mTOR, mammalian target of rapamycin; p-Akt, phosphorylated PKB; p-PI3K, phosphorylated PI3K).

Fig 4: MTNR1A inhibitor abolished the alleviative effects of NGR1 on the expressions of proteins involved in autophagy and apoptosis in HT-22 cells induced by rapamycin. After being pretreated with rapamycin (50 µg/ml) for 4 h, the cells were exposed to NGR1 (20 µM) or NGR1 (20 µM) and the MTNR1 inhibitor (10 µM) for 48 h. (A) Protein bands. (B)–(J) Gray intensity analysis of the protein bands. n = ?/group. *p < 0.05, **p < 0.01, ***p < 0.001 vs. control group. #p < 0.05, ##p < 0.01, ###p < 0.001 vs. RAPA + MTNR1 inhibitor + NGR1 group. (NGR1: notoginsenoside R1; MTNR1: melatonin receptors 1; 3-MA, 3-methyladenine; mTOR, mammalian target of rapamycin; p62, ubiquitin-binding protein p62 or sequestosome-1; p-Akt, phosphorylated PKB; p-PI3K, phosphorylated PI3K).

Fig 5: Effects of Angelica sinensis (AS) extract on phosphorylation of PI3K, AKT, and VEGFR2 in follicle microvascular endothelial-like cells of chicken (FMECs). ß-actin was used as an internal reference protein. All experiments were performed in triplicate, and the data are the mean ± S.E.M (*P < 0.05, **P < 0.01, ***P < 0.001).