Flow cytometric measurement of RNA synthesis based on bromouridine labelling and combined with measurement of DNA content or cell surface antigen.
RNA synthesis can be analysed in nuclei or cells labelled with 5-bromouridine (BrUrd) and stained using cross-reacting anti-bromodeoxyuridine (BrdUrd) antibody. Flow cytometric dual parameter analysis of BrUrd incorporation and DNA content in nuclear suspensions of human blood lymphocytes showed that RNA synthesis increased within the first 24 hours of phytohemagglutinin (PHA) stimulation, reaching a maximum at 48 hours, when cells had entered the cell cycle. Using a new method for flow cytometric dual parameter analysis of BrUrd incorporation and a cell surface antigen, spontaneous RNA synthesis in HL-60 and K-562 cells was measured simultaneous with CD13 expression.
Jensen
P O
PO
Finsen Laboratory, Rigshospitalet, Copenhagen, Denmark.
Larsen
J
J
Larsen
J K
JK
D015703:Antigens, CD / Q000032:analysis; D015214:Antigens, Differentiation, Myelomonocytic / Q000032:analysis; D000951:Antigens, Neoplasm / Q000032:analysis; D000954:Antigens, Surface / Q000032:analysis; D001976:Bromouracil / Q000031:analogs & derivatives; D018826:CD13 Antigens; D002453:Cell Cycle / Q000502:physiology; D004273:DNA, Neoplasm / Q000032:analysis; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D007938:Leukemia / Q000378:metabolism / Q000473:pathology*; D012334:RNA, Neoplasm / Q000096:biosynthesis*; D014407:Tumor Cells, Cultured; D014529:Uridine / Q000031:analogs & derivatives
D015703:Antigens, CD; D015214:Antigens, Differentiation, Myelomonocytic; D000951:Antigens, Neoplasm; D000954:Antigens, Surface; D004273:DNA, Neoplasm; D012334:RNA, Neoplasm; D001976:Bromouracil; D018826:CD13 Antigens; C006824:5-bromouridine; D014529:Uridine