Fig. 2

Characteristics of patient-derived c.del1380CDH1 SB.mhdgc-1 cancer cells. a Light microscopy image of c.del1380 CDH1 SB.mhdgc-1 cells (10× magnification). b SB.mhdgc-1 cells lack E-cadherin expression but express CEA. Immunoflurescence of SB.mhdgc-1 cells stained with anti-E-cadherin (left) and anti-CEA (right; both green), nuclei stained with DAPI (blue). Gastric cancer lines N87 and AGS (for E-cadherin), BxPC3 and HeLa cells (for CEA) shown as positive and negative controls. c Karyotype and SKY images of SB.mhdgc-1 cells show features consistent with human cancer cells including aneuploidy (such as in chromosomes 1 and 2) and chromosomal translocations (in chromosomes 3:5 and 4:8). d Light microscopy of c.del1380CDH1 SB.mhdgc-1 cancer cells grown under ultra-low attachment conditions in FBS-free media. Top (14 days of culture), three dimensional multicellular spheroid (MCS) clusters with compact, amorphous center. Bottom (33 days of culture), c.del1380CDH1 SB.mhdgc-1 spheroids have rounded up, became more compact, and formed basal membranes (arrows; 20× magnification). e Patient-derived SB.mhdgc-1 cells harbor c.1380del CDH1 germline mutation. Deep sequencing of Hg19 CDH1 locus (chr16: 68,771,195-68,869,444, NM_004360) in c.del1380CDH1 SB.mhdgc-1 spheroids (top) and SB.mhdgc-1 cells 2D monolayer cells (bottom); 40 base sequences of 14 genomic DNA fragments around c.1380 are shown. Artifacts of alignment as identified by BWA are shown in blue