pt3 ef1α myristoylated akt myrakt vector  (Addgene inc)

Journal: Journal of Cancer Research and Clinical Oncology

Article Title: AKT1-Mediated NOTCH1 phosphorylation promotes gastric cancer progression via targeted regulation of IRS-1 transcription

doi: 10.1007/s00432-024-06039-z

Figure Lengend Snippet: AKT1 Enhances Notch1 Nuclear Translocation via Phosphorylation. A : Co-IP confirms the interaction between Notch1 and AKT1. B : In vitro kinase assays reveal AKT1-induced Notch1 phosphorylation. C : A stable MGC803 cell line overexpressing AKT1 is treated ± LY294002; Western Blot assesses Flag-tagged AKT1 levels and IP measures Notch1’s RXX(S*/T*) motif phosphorylation. D : Statistical analysis of phospho-Akt substrate expression in Notch1 from C. E - H : After AKT1 overexpression and LY294002 treatment, subcellular fractionation is performed, and Western Blot analyzes Notch1 and Notch1-IC levels, with subsequent statistical analysis. I : Phosphoprotemic analysis identifies the phosphorylation site on Notch1. J : Immunofluorescence tests AKT1’s effect on Notch1 nuclear localization (×200). K : The quantification of Notch signal intensity in the nucleus. n = 3. Significance: vs. NC group, # P < 0.05, ## P < 0.01, ### P < 0.001; vs. oe-Myr-AKT1 group, * P < 0.05, *** P < 0.001

Article Snippet: The overexpression human Flag-Myr-AKT1 and Notch-IC lentiviral vectors were purchased from Shanghai Hanheng Bio-Tech Co., Ltd.

Techniques: Translocation Assay, Co-Immunoprecipitation Assay, In Vitro, Western Blot, Expressing, Over Expression, Fractionation, Immunofluorescence

Journal: Journal of Cancer Research and Clinical Oncology

Article Title: AKT1-Mediated NOTCH1 phosphorylation promotes gastric cancer progression via targeted regulation of IRS-1 transcription

doi: 10.1007/s00432-024-06039-z

Figure Lengend Snippet: AKT1-Phosphorylated Notch1 Enhances Gastric Cancer Progression via through the Regulation of IRS-1. A , B : MGC803 cells overexpressing AKT were treated with LY294002, and Western Blot confirmed AKT1’s regulation of IRS-1 expression, with semi-quantitative analysis ( n = 3 per group). Significance: ### P < 0.001 vs. oeNC; ** P < 0.01 vs. oe-Myr-AKT1. C - K : In HGC-27 and MGC803 cells with AKT1 overexpression, LY294002 and shIRS-1 lentivirus were applied to assess the effects on cell proliferation (EdU) (C-E, ×100), invasion (Transwell) ( F , H , I , ×200), and migration (scratch) (G, J, K, ×40) ( n = 3 per group). Significance: ## P < 0.01, ### P < 0.001 vs. oeNC; * P < 0.05, ** P < 0.01, *** P < 0.001 vs. oe-Myr-AKT1

Article Snippet: The overexpression human Flag-Myr-AKT1 and Notch-IC lentiviral vectors were purchased from Shanghai Hanheng Bio-Tech Co., Ltd.

Techniques: Western Blot, Expressing, Over Expression, Migration

Journal: Journal of Cancer Research and Clinical Oncology

Article Title: AKT1-Mediated NOTCH1 phosphorylation promotes gastric cancer progression via targeted regulation of IRS-1 transcription

doi: 10.1007/s00432-024-06039-z

Figure Lengend Snippet: AKT1-Mediated Notch1 Phosphorylation Promotes the Growth of Gastric Cancer Cells In Vivo through the Regulation of IRS-1. A : Nude mice were injected with MGC803 cells overexpressing AKT1 alone or co-transfected with shIRS-1, with LY294002 treatment to monitor tumorigenesis. B : Statistical analysis of tumor volume at different time points ( n = 4 per group). C - J : IHC (×200) and Western blot analyzed EMT marker expression in tumor tissues ( n = 3 per group). K - M : Western blot assessed AKT1 expression and phosphorylation, with semi-quantitative analysis ( n = 3 per group). N - Q : Expression levels of IRS-1 and Notch1 were evaluated by IHC (×200) and Western blot ( n = 3 per group). I - S : Notch1-IC expression was detected using Western blot ( n = 3 per group). Significance: vs. oeNC group, ## P < 0.01, ### P < 0.001; vs. oe-Myr-AKT1 group, * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: The overexpression human Flag-Myr-AKT1 and Notch-IC lentiviral vectors were purchased from Shanghai Hanheng Bio-Tech Co., Ltd.

Techniques: In Vivo, Injection, Transfection, Western Blot, Marker, Expressing

Journal: Journal of Cancer Research and Clinical Oncology

Article Title: AKT1-Mediated NOTCH1 phosphorylation promotes gastric cancer progression via targeted regulation of IRS-1 transcription

doi: 10.1007/s00432-024-06039-z

Figure Lengend Snippet: A mechanistic illustration of AKT1 phosphorylating notch1-IC to regulate IRS-1 expression

Article Snippet: The overexpression human Flag-Myr-AKT1 and Notch-IC lentiviral vectors were purchased from Shanghai Hanheng Bio-Tech Co., Ltd.

Techniques: Expressing

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Evaluation of a Dual PI3K/mTOR Inhibitor PF-04691502 against Bladder Cancer Cells

doi: 10.1155/2022/8110796

Figure Lengend Snippet: PF-04691502 inhibits the PI3K/Akt/mTOR pathway in bladder cancer cells. (a) T-24 and 5637 cells were treated with the indicated doses of PF-04691502 for 24 (h) and the levels of specific proteins were measured using western blot. (b) T-24 and 5637 cells were transfected with an empty vector (EV) or a vector expressing myr-Akt (OV Akt) for 12 h; then, the cells were treated with or without PF-04691502 for another 24 h and the specific proteins were measured using western blots. (c) T-24 and 5637 cells were treated as indicated above, and cellular apoptosis was measured. (d) Cellular viabilities of the cells were measured. Data are representative of at least three independent experiments ( ∗ P < 0.05; ∗∗ P < 0.01).

Article Snippet: The myr-Akt vector was purchased from Addgene (USA). siRNA against PTEN (si-PTEN) and negative scramble control (si-NC) were purchased from RioBio Ltd. (China).

Techniques: Western Blot, Transfection, Plasmid Preparation, Expressing