Figure 3

Comparison of Turtle and Padlock Probes in FFPE cells. Representative results of RNA detection with both Turtle and Padlock Probes in cells treated similarly to pathological routine specimens (fixed in formalin and embedded in paraffin). The counterstain is DAPI producing blue cell nuclei. A-F) Detection of 5S rRNA (green signals) and hTR (red signals) in FFPE Hela cells. A-C) RNA detection using Turtle Probes targeting 5S rRNA and hTR on FFPE Hela cells. A) Using a final Turtle Probe concentration of 100 nM. B) Using a final Turtle Probe concentration of 10 nM. C) Using a final Turtle Probe concentration of 1 nM. D-F) RNA detection using Padlock Probes targeting 5S rRNA and hTR. D) Using a final Padlock Probe concentration of 100 nM. E) Using a final Padlock Probe concentration of 10 nM. F) Using a final Padlock Probe concentration of 1 nM. hTR was only detected consistently with the highest concentration of the Turtle Probe. G-L) Detection of 28S rRNA (green signals) and hTR (red signals) in FFPE Hela cells. G-I) RNA detection using Turtle Probes targeting 28S rRNA and hTR on FFPE Hela cells. G) Using a final Turtle Probe concentration of 100 nM. H) Using a final Turtle Probe concentration of 10 nM. I) Using a final Turtle Probe concentration of 1 nM. J-L) RNA detection using Padlock Probes targeting 5S rRNA and hTR. J) Using a final Padlock Probe concentration of 100 nM. K) Using a final Padlock Probe concentration of 10 nM. L) Using a final Padlock Probe concentration of 1 nM. A 100× objective was used and scale bar is 50 μm.