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. Author manuscript; available in PMC: 2011 Dec 2.
Published in final edited form as: Clin Cancer Res. 2010 Aug 3;16(16):4246–4255. doi: 10.1158/1078-0432.CCR-10-1152

Fig. 3.

Fig. 3

Colocalization of miR-155 with CD45 immune cell marker. A, serial 4-μm FFPE tumor tissue sections of the indicated organs were stained with H&E to reveal histologic features or subjected to standard ISH assays using FAM2×-tagged probe against miR-155. The miR-155 signal was revealed by TSA reaction with FITC-tyramine (green). CK19 expression was revealed by TSA reaction with Dylight680-tyramine (red). After a HIER with citrate, CD45 expression was revealed by TSA reaction with rhodamine-tyramine (orange). Tissue sections were counterstained with nuclear marker 4′,6-diamidino-2-phenylindole (blue). B, line profile analysis was used to quantitate the intensity of RNA or protein expression. Background intensity was subtracted and intensity values were normalized setting the point with maximum intensity to 100, and calculating other values in relation to this reference. miRNA expression pattern was plotted as a line; independently, expression patterns of each protein were plotted as stacked areas. Displayed images were modified by optimizing the contrast with the process enhancement function from cellSens software package (Olympus), please see Supplementary Fig. S4 for raw fluorescent images and details on signal analysis.