Novel use of amine hydrochloride compound
A hydrochloride compound and compound technology, applied in the field of compounds, can solve the problems of small number of small molecular compounds and poor specificity, etc.
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[0032] (1) Screening of drugs that promote TFEB nuclear entry
[0033] ① 96-well plate plating: Collect cells, adjust the concentration of the cell suspension according to the counting results of the hemocytometer, plate to make the density of the cells to be tested 8000 / well, add 200 μL of medium to each well, and fill the edge of the 96-well plate with PBS.
[0034] ②5% CO 2 After incubation at 37° C. for 24 h, drugs (dimethyl sulfoxide (DMSO), compound I) were added at a final concentration of 5 μM.
[0035] ③ After 24 hours of dosing, take pictures with a fluorescence microscope and observe. figure 1 The results showed that compound I could significantly promote the nuclear entry of TFEB-GFP in HeLa cells stably expressing TFEB-GFP.
[0036]
[0037] (2) Nucleoplasmic separation technology
[0038]Using the cytoplasmic protein extraction reagents CERⅠ and CERⅡ, under the condition of low osmotic pressure, the cells are fully expanded, and then the cell membrane is de...
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