RNA-DNA FISH of the single mRNA molecules of EGFP and the telomeres (DNA).

<p>Telomere DNA was detected by Cy3-labeled oligonucleotide probes (shown in red). Nuclei were stained by DAPI (blue). The cells used were mouse fibroblast cells transiently transfected by an EGFP-expressing plasmid. The single mRNA molecules of EGFP were detected by a set of 20 oligonucleotides labeled by Cy5 (shown in grey). Two enlarged images of the single molecule RNA signals are shown on the right side. The images of the single molecule RNA signals are deconvolved single-layer z-sections. A scale bar of 10 µm is shown in each Dapi image. (A) RNA FISH only. (B) RNA-DNA FISH. (C) The total fluorescent signal intensity of the EGFP single mRNA molecule signals per cell (n = 50) measured from the Cy5 channel was normalized by the total EGFP fluorescent intensity per cell measured from the “green” channel. The data was presented in box-and-whisker plots. The P-value was calculated using Student t-test.