Journal: World Journal of Gastrointestinal Oncology
Article Title: Rapamycin suppresses small bowel adenocarcinoma HUTU 80 cells proliferation by inhibiting hypoxia-inducible factor-1α mediated metabolic reprogramming
doi: 10.4251/wjgo.v17.i9.109378
Figure Lengend Snippet: Effects of rapamycin on cell cycle of HUTU 80 cells. A: Flow cytometry was used to assess cell cycle changes; B: Statistical map of cell cycle distribution; C: Detection of protein expression of cell cycle proteins by western blotting; D: Statistical graph of protein expression. Data are presented as the mean ± SD ( n = 3). a P < 0.05. b P < 0.01. c P < 0.001. PI-A: Propidium iodide-area; RAPA: Rapamycin; NS: No significance; CDK2: Cyclin-dependent kinases 2.
Article Snippet: The samples were transferred to a polyvinylidene difluoride membrane, blocked with rapid blocking buffer (cat. No. P0252; Beyotime Institute of Biotechnology) for 15 minutes at 4 °C, incubated overnight with antibodies including cyclin-dependent kinases 2 (CDK2) (cat. No. 60312-1-Ig; ProteinTech Group, diluted 1:1500), mTOR (cat. No. AF6308; Affinity Biosciences, diluted 1:1500), phosphorylated (p)-mTOR (cat. No. AF3308; Affinity Biosciences, diluted 1:1500), 70 kDa ribosomal protein S6 kinase (p70S6k) (cat. No. AF6226; Affinity Biosciences, diluted 1:1500), p-p70S6k (cat. No. AF3228; Affinity Biosciences, diluted for 1:1500), eukaryotic translation initiation factor 4E-binding protein 1 (4E-BP1) (cat. No. AF6432; Affinity Biosciences, diluted 1:1500), p-4E-BP1 (cat. No. AF3830; Affinity Biosciences, diluted 1:1500), HIF-1α (cat. No. BF8002; ProteinTech Group, diluted 1:10000), and β-actin (cat. No. 66009-1-lg; ProteinTech Group, diluted 1:1500).
Techniques: Flow Cytometry, Expressing, Western Blot
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