Fig. 2

a Western blot analysis of HeLa cells over-expressing FAT1 with mAb198.3. HeLa cells were transiently transfected with pcDNA3-FAT1 plasmid and the empty vector (Mock). At 48 h post transfection HeLa cell total lysates (25 micrograms/lane, corresponding to approximately 105 cells) were loaded onto SDS-PAGE, transferred to nitrocellulose membranes and probed with mAb198.3. Molecular weight standards (MW) are reported on the left side. Arrow marks the expected protein size as determined from protein sequence. FAT1 detection in colon adenocarcinoma tissues by IHC. In contrast to normal colon tissues b, expression of FAT1 was significantly upregulated in colon adenocarcinoma tissues c-e. Bar, 50 μm. FAT1 detection in colon adenocarcinoma tissues by IHC. In contrast to non-adsorbed mAb198.3 f-g, the colon cancer tissue recognition was abolished by adsorbing the antibody on the FAT1 recombinant domain, further demonstrating the specificity of the antibody recognition h-i