Journal: The Journal of International Medical Research

Article Title: Paris polyphylla 26 triggers G2/M phase arrest and induces apoptosis in HepG2 cells via inhibition of the Akt signaling pathway

doi: 10.1177/0300060519826823

Figure Lengend Snippet: PP-26 inhibited the expression of PI3K/Akt signaling components. The expression levels of p-Akt (thr308), p-Akt (ser473), p-GSK3β (ser9), and p-FoxO3a (Ser318/321) were reduced by treatment with PP-26 for 24 h, while the expression levels of Akt, GSK3α/β, and FOXO3a did not change significantly. * P < 0.05, ** P < 0.01, ANOVA

Article Snippet: Antibodies against Akt (#9272), p-Akt (Ser473, #4060), p-Akt (Thr308, #13038), Bcl-2 (#4223), cyclin B1 (#12231), cyclin D1 (#2978), Bax (#14796), cyclin E2 (#4132), CDK4 (#12790), cdc2 (#28439), Bcl-xL (#2764), p-cdc2 (Tyr15, #4539), caspase-9 (#9502), cleaved caspase-9 (#52873), caspase-3 (#9665), cleaved caspase-3 (#9664), FOXO3a (#2497), p-FOXO3a (Ser318/321, #9465), GSK3α/β, p-GSK3β (Ser9, #5676), GAPDH (#2118), Mcl-1 (#39224), Myt-1 (#4282), PARP (#9532) and cleaved PARP (#5625) were purchased from Cell Signaling Technology Ltd. (Danvers, MA, USA).

Techniques: Expressing

Journal: The Journal of International Medical Research

Article Title: Paris polyphylla 26 triggers G2/M phase arrest and induces apoptosis in HepG2 cells via inhibition of the Akt signaling pathway

doi: 10.1177/0300060519826823

Figure Lengend Snippet: Schematic representation of the mechanism of PP26 induction of G2/M-phase arrest in HepG2 cells. The Myt1 protein kinase functions to negatively regulate Cdc2-cyclin B complexes by phosphorylating Cdc2. Akt-mediated GSK3β and FoxO3a signaling is involved in the apoptosis of cancer cells. Our study showed that PP-26 caused upregulation of p21 and Myt-1 expression, phosphorylation of cdc2 at Tyr15, and inhibition of the production of the cdc2/cyclin B complex. Inhibition of the Akt pathway is important in PP-26 mitochondrial-associated apoptosis in HepG2 cells

Article Snippet: Antibodies against Akt (#9272), p-Akt (Ser473, #4060), p-Akt (Thr308, #13038), Bcl-2 (#4223), cyclin B1 (#12231), cyclin D1 (#2978), Bax (#14796), cyclin E2 (#4132), CDK4 (#12790), cdc2 (#28439), Bcl-xL (#2764), p-cdc2 (Tyr15, #4539), caspase-9 (#9502), cleaved caspase-9 (#52873), caspase-3 (#9665), cleaved caspase-3 (#9664), FOXO3a (#2497), p-FOXO3a (Ser318/321, #9465), GSK3α/β, p-GSK3β (Ser9, #5676), GAPDH (#2118), Mcl-1 (#39224), Myt-1 (#4282), PARP (#9532) and cleaved PARP (#5625) were purchased from Cell Signaling Technology Ltd. (Danvers, MA, USA).

Techniques: Expressing, Phospho-proteomics, Inhibition

Journal: Journal of Translational Medicine

Article Title: A novel cytoprotective peptide protects mesenchymal stem cells against mitochondrial dysfunction and apoptosis induced by starvation via Nrf2/Sirt3/FoxO3a pathway

doi: 10.1186/s12967-017-1144-5

Figure Lengend Snippet: Nrf2/Sirt3/FoxO3a pathway participated in the protective mechanism of CHBP against starvation. a MSCs were treated with siRNA targeting Sirt3 and then subjected to starvation with CHBP (10 nmol/L). Total protein was prepared from cultured cells. Expressions of Nrf2, Sirt3, total FoxO3a and p-FoxO3a were analyzed by western blotting. b Sirt3 siRNA treated MSCs were subjected to serum deprivation and apoptosis was detected by Annexin V/PI asaay. Experiments were performed in triplicate. Data are expressed as mean ± standard deviation. * P < 0.05, ** P < 0.01

Article Snippet: Briefly, total proteins from MSCs were separated on SDS–polyacrylamide gels, transferred onto nitrocellulose membranes, blocked and incubated with anti-Nrf2, anti-Sirt3, anti-total FoxO3a, and phosphorylated-FoxO3a (p-FoxO3a, Ser253 and Ser318/321) antibodies (1:1000, Cell Signaling Technology) and anti-tubulin antibodies (1:10000, Abcam) overnight at 4 °C.

Techniques: Cell Culture, Western Blot, Standard Deviation