Brm targeting compounds and associated methods of use
A compound and chemical structure technology, applied in drug combination, organic chemistry, pharmaceutical formulation, etc., can solve the problem of SMARCA2 hindering the development of effective therapeutic agents
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[2496] Assay and Degradation Data
[2497] Western blot screen for BRM degradation in SW1573 cells
[2498] To assess BRM degradation (D max and DC 50 ), the cells were seeded in 96-well black / transparent bottom plate at 8000 / well, and each well was 180L DMEM growth medium (containing 1% Pen-Strep, 1% HEPES and 10% FBS). Plates were incubated overnight to allow adhesion. The next morning, cells were treated by adding 20L of 10X compound concentration of interest (1% DMSO) to the appropriate wells and returned to the incubator overnight (18-20 hours). The final concentration of DMSO was 0.1%.
[2499] For lysis, adhered cells were washed once with 100 L DPBS. Cells were lysed in 40L 1X RIPA + HALT protease inhibitors on ice for 10 minutes and then frozen until use at -80C. Thawed lysates were cleaned by filtration in 1.2m filter plates, or by spinning at 2300g for 30 min at 4C.
[2500] For blots, for each Western sample, 30 L of lysate was added to 10 L of 4X LDS sample...
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