Detection of living cells that express AP1 using a fluorolabeled DNA probe.
Activator protein 1 (AP1) is a complex of Fos and Jun, and it regulates the transcription of genes possessing the AP1-binding sequence. The purpose of this study was to detect living cells that express AP1 after stimulation with a tumor promoter. The Fos and Jun components of AP1 were induced rapidly and transiently in PC12 cells following the addition of phorbol ester (phorbol 12-myristate 13-acetate, PMA). The DNA fragment containing the AP1-binding sequence was combined with ethidium bromide, which was used as a fluorescent probe. The probe was transfected into the cells using cationic liposomes. Fluorescence in the transfected cells was observed using a fluorescence microscope. The nuclei of transfected cells emitted strong fluorescence in the presence of PMA, whereas weak fluorescence was retained in the cytoplasm in its absence. The former phenomenon is evidence that AP1 combined with the fluorescent probe was transported into the nuclei. This study suggests that such a fluorolabeling method is feasible to detect living AP1-expressed neurons.
Shimokawa
N
N
Department of Physiology 1st Division, Gunma University School of Medicine, Maebashi City, Japan.
Miura
M
M
D000818:Animals; D002467:Cell Nucleus / Q000737:chemistry; D003593:Cytoplasm / Q000737:chemistry; D015342:DNA Probes; D004996:Ethidium; D005456:Fluorescent Dyes; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D008081:Liposomes; D016716:PC12 Cells / Q000737:chemistry; D016760:Proto-Oncogene Proteins c-fos / Q000032:analysis / Q000235:genetics; D016755:Proto-Oncogene Proteins c-jun / Q000032:analysis / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis; D012334:RNA, Neoplasm / Q000096:biosynthesis; D051381:Rats; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D018808:Transcription Factor AP-1 / Q000032:analysis*; D014162:Transfection
D015342:DNA Probes; D005456:Fluorescent Dyes; D008081:Liposomes; D016760:Proto-Oncogene Proteins c-fos; D016755:Proto-Oncogene Proteins c-jun; D012333:RNA, Messenger; D012334:RNA, Neoplasm; D018808:Transcription Factor AP-1; D004996:Ethidium; D013755:Tetradecanoylphorbol Acetate