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Stem Cells 36, 337–348. 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 2017

Kamarudin, T.A., Bojic, S., Collin, J., Yu, M., Alharthi, S., Buck, H., Shortt, A., Armstrong, L., Figueiredo, F.C., Lako, M.

Notes: RNA was extracted from the cells collected from differentiating hESC and hiPSC at days 0, 9, and 20 using the ReliaPrep RNA Cell Miniprep System. The RNA quality was evaluated using the Nanodrop 2000 spectrophotometer (Thermo Fisher Scientific, MA). One microgram of extracted RNA was converted into cDNA using reverse transcription (GoScript Transcription System). Quantitative real‐time polymerase chain reaction (qRT‐PCR) was carried out using the QuantStudio 7 Flex Real‐Time PCR System (Thermo Fisher Scientific, MA) and GoTaq qPCR Master Mix. pGL3 BRE Luciferase was used for plasmid lipofection to transfect the cells in each well of a 24-well plate. Cells that were transfected with empty vector (pGL3‐Basic) or BMP reporter (pGL3 BRE Luciferase) were cotransfected with empty Renilla vector (pRL‐Null). Luciferase activities were evaluated with a Dual‐Luciferase Assay System. (4982)

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