AI Article Synopsis
- The study aimed to investigate how overexpressing the protein KLF4 affects cell death (apoptosis) in macrophages when subjected to heat stress.
- Researchers inserted the KLF4 gene into a vector and introduced it into Raw264.7 macrophages, then analyzed the cells after exposing them to heat stress.
- Results showed that macrophages with higher KLF4 levels had significantly increased apoptosis after heat stress, indicated by various assays confirming cell death and DNA fragmentation.
Article Abstract
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.
Conclusion: KLF4 overexpression can increase heat stress-induced apoptosis of Raw264.7 macrophages.
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