Journal: Journal of Functional Foods

Article Title: Eicosapentaenoic acid and docosahexaenoic acid suppress colonic tumorigenesis in obese mice

doi: 10.1016/j.jff.2024.106164

Figure Lengend Snippet: Fig. 7. Schematic diagram of the mechanism of EPA and DHA inhibiting colon tumor in obese mice. This mechanism involves solidifying the intestinal mucosal barrier by improving the intestinal flora dysbiosis, suppressing inflammatory signaling pathways such as TNF-α/NF-κB, IL-6/STAT3, and NLRP3/IL-1β pathways, and blocking the GSK3β/β-catenin/C-myc carcinogenic pathway.

Article Snippet: Primary antibodies against NLRP3 (15101S, 1:1000), Stat3 (12640S, 1:1000), p-Stat3Tyr705 (9131S, 1:1000), ERK (9102S, 1:1000), p-ERK (4370S, 1:2000), Akt (9272S, 1:1000), p-AktSer473 (9271S, 1:1000), p-GSK3βSer9 (9336S, 1:1000), β-catenin (9562S, 1:1000), C-myc (9402S, 1:1000), Cyclin-D1 (2922S, 1:1000), PCNA (13110S, 1:1000), NF-κB (8242S, 1:1000), and horseradish peroxidase (HRP)-conjugated secondary antibodies (7074S, 1:2000) were obtained from Cell Signaling Technology, Inc. (Danvers, USA).

Techniques: Protein-Protein interactions, Blocking Assay

Journal: The International Journal of Neuropsychopharmacology

Article Title: Activation of signaling pathways downstream of the brain-derived neurotrophic factor receptor, TrkB, in the rat brain by vagal nerve stimulation and antidepressant drugs

doi: 10.1017/s1461145713000977

Figure Lengend Snippet: Fig. 6. Western blot analysis of hippocampus CREB (≈50 kDa) phosphorylation at S133 after chronic 14 d VNS, sertraline (7.5 mg/ kg/d i.p.) or DMI (10 mg/kg/d i.p.). *p<0.01, Student’s t-test. The number of rats in each group is shown in the bars.

Article Snippet: Membranes were incubated at 4 °C overnight with the following primary antibodies: anti-pY515 (1:1000 in 2.5% BSA in TBST, Abcam, USA), anti-pY705 TrkB (1:1000 in 2.5% BSA in TBST, Abcam, USA), anti-pY816 TrkB (1:4000 in 2.5% BSA in TBST, Abcam, USA), and anti-TrkB (full length, 1:10000 in 2.5% BSA in TBST, Neuromics, USA), anti-PLCγ1 (1:1000), anti-PLC γ1 Y783 (1:250 in 5% BSA in TBST, Cell Signaling Technology, Inc, USA), anti-pERK T202/Y204 (1:1000 in 5% BSA in TBST, Cell Signaling Technology, Inc, USA), anti-ERK (1:2000 in 5% BSA in TBST, Cell Signaling Technology, Inc, USA), anti-AKT (1:1000 in 5% BSA in TBST, Cell Signaling Technology, Inc, USA), anti-pAKT T308 (1:1000 in 5% BSA in TBST, Cell Signaling Technology, Inc, USA), anti-CREB (1:2000 in 5% BSA in TBST, Cell Signaling Technology, Inc, USA), anti-pCREB S133 (1:1000 in 5% BSA in TBST, Cell Signaling Technology, Inc, USA), anti- pp70 S6 kinase T389 (1:1000 in 5% BSA in TBST, Cell Signaling Technology, Inc, USA) and anti-p70 S6 kinase (1:1000 in 5% BSA in TBST, Cell Signaling Technology, Inc, USA).

Techniques: Western Blot, Phospho-proteomics

Journal: Journal of Medicinal Chemistry

Article Title: Synthetic Lethality in Pancreatic Cancer: Discovery of a New RAD51-BRCA2 Small Molecule Disruptor That Inhibits Homologous Recombination and Synergizes with Olaparib

doi: 10.1021/acs.jmedchem.9b01526

Figure Lengend Snippet: Figure 2. (A) RAD51−BRCA2 BRC repeat complex (PDB code: 1N0W). RAD51 is represented as

Article Snippet: Samples were incubated in 10% Bovine Serum Albumin (BSA) in PBS for 30 min at 37 °C and subsequently exposed to an anti-RAD51 mouse monoclonal antibody (Santa Cruz Biotechnology, 1:1000 in 5% BSA / PBS), overnight at 4 °C.

Techniques:

Journal: Cancer cell international

Article Title: GNF-7, a novel FLT3 inhibitor, overcomes drug resistance for the treatment of FLT3‑ITD acute myeloid leukemia.

doi: 10.1186/s12935-023-03142-y

Figure Lengend Snippet: Fig. 1 GNF-7 potently inhibited the proliferation of FLT3-ITD AML cells and targeted FLT3-ITD downstream signaling pathways. A AML cell line MOLM-13, MV4-11, U937 and THP-1 were treated with DMSO or increasing concentrations of GNF-7 for 48 h and the normalized cell proliferation was measured by CellTiter Glo assay. B Mononuclear cells isolated from umbilical cord blood (Normal #1, Normal #2), bone marrow of diagnosed with AML (AML #1, AML #2) and AML harboring FLT3-ITD mutation (AML #3, AML #4, AML #5) were treated with different concentrations of GNF-7 for 48 h, and normalized cell proliferation was detected by CellTiter Glo assay. C Primary bone marrow cells isolated from AML #3 and AML #4 were exposed to GNF-7 for 4 h and then detected by western blotting with antibodies against phosphorylated and total of FLT3, Stat5, AKT and ERK, respectively. D Phosphorylated and total of FLT3, Stat5, AKT and ERK in MOLM-13 and MV4-11 cells treated with GNF-7 for 4 h were detected by western blotting. E Dose response curve of GNF-7 on Ba/F3 FLT3-ITD cells in the presence or absence of IL-3. F Ba/F3 FLT3-ITD cells were treated with the different concentration of GNF-7 for 4 h and subjected to western blotting with the indicated antibodies. All experiments were repeated three times with the same results. Data are presented as mean ± SD, and P values were calculated using Student t test. *p < 0.05, ** p < 0.01, and *** p < 0.001

Article Snippet: The membrane was then blotted with specific primary antibodies against p-FLT3 (Tyr591, #3466, CST), p-AKT (Ser473, #4060S, CST), ERK (#4695S, CST), p-ERK (T202/Y204, #4370S, CST), p-Stat5 (Y694, #AP0887, Abclonal), FLT3 (#ab245116, Abcam), and Stat5 (#13179-1-AP, Proteintech), AKT (#10176-2-AP, Proteintech), actinHRP (#HRP-60008, Proteintech).

Techniques: Protein-Protein interactions, Glo Assay, Isolation, Mutagenesis, Western Blot, Concentration Assay

Journal: Cancer cell international

Article Title: GNF-7, a novel FLT3 inhibitor, overcomes drug resistance for the treatment of FLT3‑ITD acute myeloid leukemia.

doi: 10.1186/s12935-023-03142-y

Figure Lengend Snippet: Fig. 2 GNF-7 interacts with FLT3 protein. A Docking model of FLT3 bound to GNF-7: the yellow dotted line represents the hydrogen bond interaction, the green line represents the amino acid that forms hydrogen bonds with GNF-7, the cartoon represents the FLT3 protein, and the purple stick represents the GNF-7 molecule. B 2D interaction diagram of the FLT3/GNF-7 complex: GNF-7 is bound to the FLT3 protein in a pocket surrounded by LYS706, TYR696, GLY697, CYS694, ALA642, LEU818, PHE830, VAL624, LEU616, CYS695, and SER705 amino acids, GNF-7 forms hydrogen bonds with SER705 and forms hydrophobic interaction with LYS706, TYR696, GLY697, CYS694, ALA642, LEU818, PHE830, VAL624, LEU616, CYS695. C, E Thermal stabilization of FLT3 in Ba/F3 FLT3-ITD cells treated with GNF-7 (1 μM) or DMSO in various temperatures (C) and treated with various concentrations of GNF-7 (E) was analyzed through CETSA assay. D–F The density of the FLT3 bands were quantified by quantity one software. All experiments were repeated three times with the same results. Data are presented as mean ± SD, and P values were calculated using Student t test. * p < 0.05, ** p < 0.01, and *** p < 0.001

Article Snippet: The membrane was then blotted with specific primary antibodies against p-FLT3 (Tyr591, #3466, CST), p-AKT (Ser473, #4060S, CST), ERK (#4695S, CST), p-ERK (T202/Y204, #4370S, CST), p-Stat5 (Y694, #AP0887, Abclonal), FLT3 (#ab245116, Abcam), and Stat5 (#13179-1-AP, Proteintech), AKT (#10176-2-AP, Proteintech), actinHRP (#HRP-60008, Proteintech).

Techniques: Software

Journal: Cancer cell international

Article Title: GNF-7, a novel FLT3 inhibitor, overcomes drug resistance for the treatment of FLT3‑ITD acute myeloid leukemia.

doi: 10.1186/s12935-023-03142-y

Figure Lengend Snippet: Fig. 3 GNF-7 showed significant therapy effect on the mice model engrafted with Ba/F3 FLT3-ITD cells. A The mice engrafted with Ba/F3 FLT3-ITD cells were treated with vehicle, AC220 (10 mg/kg), gilteritinib (30 mg/kg) and GNF-7 (15 mg/kg) for 8 days and the percentages of leukemia cells infiltrated in peripheral blood were then analyzed by flow cytometry. B The mice engrafted with Ba/F3 FLT3-ITD cells were treated with vehicle, AC220 (10 mg/kg), gilteritinib (30 mg/kg) and GNF-7 (15 mg/kg) for 9 days and the percentages of leukemia cells infiltrated in bone marrow were then analyzed by flow cytometry. C The spleen weight in each group were measured. D The survival curve of mice was calculated. Data are presented as mean ± SD, and P values were calculated using Student t test. * p < 0.05, ** p < 0.01, and *** p < 0.001

Article Snippet: The membrane was then blotted with specific primary antibodies against p-FLT3 (Tyr591, #3466, CST), p-AKT (Ser473, #4060S, CST), ERK (#4695S, CST), p-ERK (T202/Y204, #4370S, CST), p-Stat5 (Y694, #AP0887, Abclonal), FLT3 (#ab245116, Abcam), and Stat5 (#13179-1-AP, Proteintech), AKT (#10176-2-AP, Proteintech), actinHRP (#HRP-60008, Proteintech).

Techniques: Flow Cytometry

Journal: Cancer cell international

Article Title: GNF-7, a novel FLT3 inhibitor, overcomes drug resistance for the treatment of FLT3‑ITD acute myeloid leukemia.

doi: 10.1186/s12935-023-03142-y

Figure Lengend Snippet: Fig. 4 Activity of GNF-7 against drug resistant Ba/F3 FLT3-ITD/F691L cells. A Relative proliferation of Ba/F3 populations stably expressing FLT3-ITD mutant isoforms after 48 h in various concentrations of GNF-7 were measured by CellTiter Glo assay. B IC50 values of Ba/F3 stably expressing FLT3-ITD and FLT3-ITD/F691L cells treated with various concentrations of AC220, gilteritinib and GNF-7 were analyzed by CellTiter Glo assay. C After treated with vehicle, AC220 (10 mg/kg), gilteritinib (30 mg/kg) and GNF-7 (15 mg/kg) for 8 days, the percentages of leukemia cells infiltrated in peripheral blood of mice (n = 6) engrafted with Ba/F3 FLT3-ITD/F691L cells were evaluated by flow cytometry. D After treated with vehicle, AC220 (10 mg/kg), gilteritinib (30 mg/kg) and GNF-7 (15 mg/kg) for 9 days, the burden of leukemia cells in bone marrow of mice (n = 3) which was randomly selected from each group were detected. E The spleen weights were analyzed. F The Kaplan–Meier survival curves of animal survival of mice treated with vehicle, AC220 (10 mg/kg), gilteritinib (30 mg/kg) and GNF-7 (15 mg/kg). All cell line experiments were repeated three times with the same results. P values were calculated by log-rank test and shown. Data are presented as mean ± SD, and P values were calculated using Student t test. * p < 0.05, ** p < 0.01, and *** p < 0.001

Article Snippet: The membrane was then blotted with specific primary antibodies against p-FLT3 (Tyr591, #3466, CST), p-AKT (Ser473, #4060S, CST), ERK (#4695S, CST), p-ERK (T202/Y204, #4370S, CST), p-Stat5 (Y694, #AP0887, Abclonal), FLT3 (#ab245116, Abcam), and Stat5 (#13179-1-AP, Proteintech), AKT (#10176-2-AP, Proteintech), actinHRP (#HRP-60008, Proteintech).

Techniques: Activity Assay, Stable Transfection, Expressing, Mutagenesis, Glo Assay, Flow Cytometry

Journal: Cancer cell international

Article Title: GNF-7, a novel FLT3 inhibitor, overcomes drug resistance for the treatment of FLT3‑ITD acute myeloid leukemia.

doi: 10.1186/s12935-023-03142-y

Figure Lengend Snippet: Fig. 5 GNF-7 exerts potent therapy effect on AML PDX model. A, B Primary patient cells from AML #3 or AML #4 carrying with FLT3-ITD were transplanted into busulfan pretreated NOG mice and then randomly divided into three groups. Peripheral blood of 5 mice administrated with vehicle, 15 mg/kg GNF-7 and 30 mg/kg Gilteritinib were collected at the indicated time, and the leukemia cells content was detected by flow cytometry using human CD45 antibody. C, D The percentages of leukemia stem and progenitor cells in peripheral blood were detected by flow cytometry using human CD45 and CD34 antibodies. E, F The Kaplan–Meier survival curves of animal survival of mice treated with vehicle, AC220 (10 mg/kg), gilteritinib (30 mg/kg) and GNF-7 (15 mg/kg)

Article Snippet: The membrane was then blotted with specific primary antibodies against p-FLT3 (Tyr591, #3466, CST), p-AKT (Ser473, #4060S, CST), ERK (#4695S, CST), p-ERK (T202/Y204, #4370S, CST), p-Stat5 (Y694, #AP0887, Abclonal), FLT3 (#ab245116, Abcam), and Stat5 (#13179-1-AP, Proteintech), AKT (#10176-2-AP, Proteintech), actinHRP (#HRP-60008, Proteintech).

Techniques: Flow Cytometry