[Effect of Krüppel-like factor 4 overexpression on heat stress-induced apoptosis of Raw264.7 macrophages].
OBJECTIVE
To observe the effect of Krüppel-like factor 4 (KLF4) overexpression on heat stress-induced apoptosis of Raw264.7 macrophages.


METHODS
The fragment containing full length mouse KLF4 cDNA coding sequence was inserted into the pcDNA3.1 vector and Raw264.7 macrophages were transfected with pcDNA3.1-KLF4 plasmids using lipofectamine.The expression of KLF4 was examined by Western blot in the Raw264.7 macrophages stably transfected with pcDNA3.1- KLF4 plasmids. Raw264.7 cells transfected with pcDNA3.1 and pcDNA3.1-KLF4 were exposed to heat stress (42 degrees C) for 1h and recovered at 37 degrees C for 12h. Flow cytometry, Hoechest 33258 staining assay, and DNA ladder assays were performed to assess the apoptosis.


RESULTS
The KLF4 overexpressed Raw264.7 macrophages were established. After the heat stress, flow cytometry showed that apoptotic cells increased significantly in KLF4 overexpressed cells compared with the vector control; Hoechest 33258 staining was characterized with classical changes including apoptotic body, and nuclear condensation. Agarose gel electrophoresis showed that "DNA ladder" could be observed clearly.


CONCLUSIONS
KLF4 overexpression can increase heat stress-induced apoptosis of Raw264.7 macrophages.
Liu
Mei-dong
MD
Department of Pathophysiology, Xiangya School of Medicine, Central South University, Changsha 410078, China.
Liu
Ying
Y
Liu
Jun-wen
JW
Zhang
Hua-li
HL
Xiao
Xian-zhong
XZ
D000818:Animals; D017209:Apoptosis / Q000235:genetics*; D002460:Cell Line; D005822:Genetic Vectors; D018869:Heat-Shock Response; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors / Q000235:genetics*; D008264:Macrophages / Q000166:cytology*; D051379:Mice; D010957:Plasmids; D014162:Transfection
C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors