Whole-mount in situ hybridization was performed with digoxigenin (DIG)-labeled probes as described by [9] (link). Specific antisense probes for both CD133 and VEGFR-2 were synthesized from PCR products using CD133 and VEGFR-2 clones coding for 873 bp and 348 bp regions of the respective genes. TSA-plus detection (PerkinElmer, NEL753001KT) with Cyanine 3 (CD133) and Fluorescein (VEGFR-2) substrates were used in our analysis.
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