Panels A and B show the cytoplasmic detection of miR-127 and miR-154, respectively. Panel C shows the nuclear expression of the positive control U6 small RNA as visualized using the miRCURY LNA U6 control probe. No signal was detected when cells were hybridized with the miRCURY LNA scramble probe as a negative control, as seen in panel D. Data used with kind permission from Dr. Silvana Debernardi, Institute of Cancer, London, UK. Original figure appears in Dixon-McIver A, East P, Mein CA, Cazier J-B, Molloy G, et al. (2008) Distinctive Patterns of miRNA Expression Associated with Karyotype in Acute Myeloid Leukaemia. PLoS ONE 3(5): e2141. doi:10.1371/journal.pone.0002141.
Specific detection of miR-122a (top), miR-206 (middle) and miR-124a (bottom) using miRCURY LNA miRNA Detection Probes for in situ hybridization of whole-mount zebrafish embryos. This image is a part of a complete catalog of images showing the temporal and spatial expression patterns of 115 conserved miRNAs in zebrafish embryos. The image was kindly provided by Dr. Ronald Plasterk, Hubrecht Laboratory, The Netherlands.
Specific detection of mir-206 in Gallus gallus embryo using a miRCURY LNA miRNA Detection Probe. mir-206 is expressed in all skeletal muscle cells, appearing at the onset of myogenic cell differentiation. At the pictured stage in embryonic development, mir-206 is detected in the myotomal muscle cells (Ason et al. 2006).
hsa-miR-21 was detected in tissue sections using a miRCURY LNA miRNA Detection Probe with a double DIG (5’ and 3’) label at 40 nM (A) or a single 3’ DIG label at 80 nM (B). The double DIG probe gives a more intense signal with lower background, even at a lower probe concentration.
The examples show miRCURY LNA miRNA Detection Probes with double DIG label and double FAM label for miR-124 in human normal brain samples, and miR-126 and scramble negative control in human colon cancer samples. The miR-126 and scramble probes show the same area on serial sections. Probes were incubated at 30 nM probe concentration and hybridized at 57ºC. The double FAM-labeled LNA probes show the same signal-to-noise as the double DIG-labeled LNA probes, and can be used independently or in combination to co-localize miRNAs using different hapten-labeled probes or antibodies, in the case of protein co-detection by immunohistochemistry. Images kindly provided by Boye Schnack Nielsen, Manager, Molecular Histology at Bioneer A/S, Hørsholm, Denmark.
Co-localization of miR-205 (pink) and cytokeratin (green) in ductal carcinoma in situ (DCIS). miR-205 was detected using a double fluorescein (FAM)-labeled miRCURY LNA miRNA Detection Probe, and cytokeratin was detected using a chromogenic secondary antibody. Counterstain: DAPI (blue). Images kindly provided by Boye Schnack Nielsen, Manager, Molecular Histology at Bioneer A/S, Hørsholm, Denmark.
Co-localization of miR-205 (pink) and cytokeratin (green) in the normal mammary gland. miR-205 was detected using a double fluorescein (FAM)-labeled miRCURY LNA miRNA Detection Probe, and cytokeratin was detected using a chromogenic secondary antibody. Counterstain: DAPI (blue). Images kindly provided by Boye Schnack Nielsen, Manager, Molecular Histology at Bioneer A/S, Hørsholm, Denmark.
Two duplicate dilution series of A. thaliana total RNA were hybridized with 32P-labeled DNA and miRCURY LNA miRNA Detection Probes, respectively, for A. thaliana miR-171 and miR-319. From Válóczi et al. 2004, Nucleic Acids Res. e175; reprinted with permission from Oxford University Press.
The specificity of the probe was assessed using 32P-labeled perfect match, double mismatch and single mismatch miRCURY LNA miRNA Detection Probes for the detection of miR-171 in A. thaliana flowers (1) and leaves (2). The filters were washed at either low stringency or high stringency. From Válóczi et al. 2004, Nucleic Acids Res. e175; reprinted with permission from Oxford University Press.
