Lipofectamine 3000 reagent (Thermo Fisher, Waltham, MA) was used for BMP reporter plasmid (Addgene, MA) transfection. Cells were disassociated by incubating with EDTA (0.02%) for 5 minutes. The disassociated cells were collected and centrifuged at 500g for 5 minutes. The cell pellet was resuspended into 2 ml of media and cell count was performed before replating cells at the density of 1.3 × 105 cells into one well of a BD Matrigel coated 24 well plate one day before lipofection. For plasmid lipofection, 500 ng of pGL3‐Basic or pGL3 BRE Luciferase (Promega, Madison, WI) were used to transfect the cells in each well of 24‐well plate following manufacturer's recommendations. Cells that were transfected with empty vector (pGL3‐Basic) or BMP reporter (pGL3 BRE Luciferase) were cotransfected with empty Renilla vector (pRL‐Null) (Promega, Madison, WI).
Kamarudin T.A., Bojic S., Collin J., Yu M., Alharthi S., Buck H., Shortt A., Armstrong L., Figueiredo F.C, & Lako M. (2017). Differences in the Activity of Endogenous Bone Morphogenetic Protein Signaling Impact on the Ability of Induced Pluripotent Stem Cells to Differentiate to Corneal Epithelial‐Like Cells. Stem Cells (Dayton, Ohio), 36(3), 337-348.
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