Application of FoxO1 gene in drug preparation
A technology of 1.foxo1 and 2.foxo1, applied in the field of biomedicine, can solve problems affecting quality of life, delayed fracture healing, and increased fracture rate
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[0018] 1. FoxO1 gene knockout on osteoblasts
[0019] FoxO1 gene knockout on osteoblasts was completed by using pcDNA3.1-Fox01 vector construction, interference vector construction and screening, interference with lentiviral vector construction, interference with lentiviral packaging, and virus invasion and infection of cells.
[0020] After the lentivirus infects the 3T3-E1 cells, the observation results of the stable transfection strains under the fluorescence microscope are as follows: figure 1 , the irrelevant shRNA transfection control group (marked as NC) and FoxO1-shRNA lentivirus transfected group (marked as FoxO1-shRNA) cells had significant fluorescence expression.
[0021] 2. Osteoblast culture and induced differentiation
[0022] 1) Culture of osteoblasts
[0023] Prepare α-MEM culture solution containing 10% FBS, 1% antibiotic and 1ug / ml blasticidin, and place it in a 37°C water bath to preheat for 20 minutes; normal sugar control group (NG), high sugar group (H...
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