Figure 2.

Validation of fusion genes in canine HSA. A, MYO16-PTK2 fusion gene track and visualization of the breakpoint in UCSC Genome Browser (Canfam3.1). B, Sanger sequencing result of the PCR product for MYO16-PTK2 fusion gene. C, Schematic illustration of the putative MYO16-PTK2 fusion gene and designed FISH probes. D, Detection of the MYO16-PTK2 fusion gene on primary canine HSA tissue by FISH. In wild type cells an association between BAC clones 183H20 (red) and 385H13 (green) can be appreciated, both showing independent localization from 451H13 (aqua). A portion of tumor cells shows a breakage within 451H13, with one half of that signal associating with 183H20, and independent of the localization of 385H13 indicating the existence of the MYO16-PTK2 fusion at the genomic level. E, Arrows indicate the amplification of the MYO16-PTK2 fusion gene. F, Detection of the GABRA3-FLT1 fusion gene by FISH. The GABRA3-FLT1 fusion is identified by break-apart FISH probes for proximal FLT1 (clone 363B20; red) at CFA 25 and distal FLT1 at CFA 25 (clone 235H9; green). Split FLT1 genes indicate the fusion event identified by single color signal (white arrows). Dual colors represent the intact FLT1 gene (grey arrows). G, DNA amplification of FLT1 gene.