Fig 4.

Calcineurin and NFAT are necessary for constitutive and En-1 inducible CryAB gene transcription. HL-1 cells were transfected either with CryAB-Luc (A), or with mutBE4-Luc (B). Reporter constructs were cotransfected with an expression plasmid encoding either cain or pECE-flag (parental vector) (A), or either dominant negative NFAT (dnNFAT) plasmid or pcDNA3. (parental vector) (B). Cells were treated with 50 nM En-1 or vehicle (water). Renilla luciferase expression was used for normalization. Luciferase activities in cells cotransfected with cain or dnNFAT are expressed relative to the mean value obtained from cells cotransfected with parental vector and treated with the vehicle only. Cells cotransfected with cain or dnNFAT and exposed to En-1 versus cotransfected with parental vector exposed to En-1. Experiments have been repeated at least three times. Data are mean ± SE. *P < 0.05.