Simultaneous detection of double-stranded RNA-induced protein kinase and its specific mRNA by single cell analysis.
Fluorescent in situ hybridization was combined with flow cytometry to detect the expression of the double-stranded-RNA-induced protein kinase (PKR) in single cells. Labeled anti-sense oligonucleotide was used to target the specific mRNA while the protein was targeted with an antibody. It was demonstrated that the PKR-mRNA signal could be protected through a lengthy immunostaining procedure. The expression pattern of the PKR-mRNA with respect to DNA content was shown to be comparable to that of 18S ribosomal RNA.
Lozano
Mónica M
MM
Chemical Engineering Program, University of California La Jolla, San Diego, 92093, USA.
Chantawansri
Tanya L
TL
Chau
Pao C
PC
D045744:Cell Line, Tumor; D005434:Flow Cytometry / Q000379:methods; D005456:Fluorescent Dyes; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods; D011494:Protein Kinases / Q000235:genetics* / Q000378:metabolism; D012330:RNA, Double-Stranded / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D015203:Reproducibility of Results
D005456:Fluorescent Dyes; D012330:RNA, Double-Stranded; D012333:RNA, Messenger; D011494:Protein Kinases