Figure 5. KLF4 transient transfection in HCE represses endoglin and ODC gene expression

A: Detection of the messengers encoding KLF4, endoglin and ODC by RT-PCR on total mRNA extracted from human total cornea. B: HCE cells were transfected with 2 μg of pMT3 or 2 μg of pMT3-KLF4 vector and 2 μg of pCMV-β-galactosidase. After 36 h transient transfection, total mRNA was extracted and submitted to RT-PCR experiments using oligonucletide primers specific of KLF4, endoglin, ODC and GAPDH. Fold induction or reduction of KLF4, endoglin and ODC mRNA levels were normalized using GAPDH mRNA levels. Determination of the β-galactosidase activity was realized to rationalize with the transfection percentage efficiency. The data presented were obtained from five independent cultures and PCR amplifications. They are shown with their mean ± standard deviations of five independent assays. KLF4 was normalized with the efficiency percentage based on the simultaneous determination of β-galactosidase. ODC and endoglin levels and variations are similarly reported normalized to β-galactosidase enzymatic activity.