Zhang Jun, Ma Zeng-yi, Wang Yue-ling, Wang Ji-yan, Hu Yan-zhong
Key Laboratory of Cell and Molecular Immunology, Medical College of Henan University, Kaifeng 475000, China.
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Zhonghua Yan Ke Za Zhi. 2013 Nov;49(11):1029-31.
To establish mouse lens epithelial cell lines with the genotype of Hsf4-/-.
The expended mouse lens epithelial cells, which were generated from P6 Hsf4-deficient mouse lens epithelia, were immortalized with SV40-T-antigen and named MLEC/Hsf4-/- cell. The expression of alpha A-crystallin was immunoblotted. Hsf4b cDNA was reconstituted by transiently transfection.
The SV40-immortalized cells were in adherent growth mode with spindle morphology, pseudopodia, clear nuclear boundary membrane and cytoplasm translucent. Immunoblotting results indicated that the lens biomarker protein alpha A-crystallin was expressed in MLEC/Hsf4-/- cells. Reconstitution of Hsf4b into MLEC/Hsf4-/- cells upregulated the expression of Hsp25.
The SV40-immortalized MLEC/Hsf4-/- cells have the lens epithelial characteristics and could be used as a tool for studying the signal transduction in vitro.