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Generally, 2500 cells were plated in each well of a 384-well plate with 100 μL of medium, and 500 nL of the 1 mM compound solutions were added. After incubating for 4 days, cells were lysed, and ALP activities were assayed (see Supporting Information for details).
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13
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0011948235
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note
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(13) The promoter region of Cbfa1/Runx2 gene (1.8 kb) was cloned and inserted directly upstream of the luciferase gene. Cells were transfected with reporter plamid and plated in 96-well plates. After incubating for 4 days, luciferase activity was assayed using Bright-Glo Luciferase reagent (see Supporting Information for details).
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0011917664
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Cells were fixed with 10% formalin solution and stained with an ALP staining kit from Sigma Diagnostics (see Supporting Information for details).
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