FECD corneal endothelial cell lines (F35T and F45SV) and a control corneal endothelial cells (W4056–17-001579) were cultured as described (26 (link)) The C9 and VVM84 skin fibroblasts were maintained at 37°C and 5% CO2 in Minimal Essential Media Eagle (MEM) (Sigma, M4655) supplemented with 15% heat inactivated fetal bovine serum (Sigma) and 0.5% MEM nonessential amino acids (Sigma).
Endothelial or fibroblast cells were seeded in 6-well plate at 90% confluence. At the next day, actinomycin D was added into the wells at 5 μg/ml final concentrations. Cells were harvest using TRIzol agent (Sigma) at different time point. The TCF4 RNA levels were analyzed by qPCR.
Endothelial or fibroblast cells were seeded in 6-well plate at 90% confluence. At the next day, actinomycin D was added into the wells at 5 μg/ml final concentrations. Cells were harvest using TRIzol agent (Sigma) at different time point. The TCF4 RNA levels were analyzed by qPCR.
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