Fig 1: Long-term spheroidogenesis of WT and CDH1 KO HCT116 cells. (A) When spheroidogenesis was maintained for 21 days, the number of spheroids was decreased while sizes of spheroids were increased in a time-dependent manner in both cells. Interestingly, some spheroids from CDH1 KO cell line lost their morphological integrity (arrows). Scale bar=500 µm. (B) Immunocytochemistry (magnification, ×4 and ×20) using with an anti-E-cadherin antibody for spheroids revealing decreased E-cadherin in CDH1 KO cells. Arrows indicate necrosis. *P<0.05, ***P<0.001 vs. Day 7. WT, wild-type; KO, knock-out.
Fig 2: General characteristics of WT and CDH1 KO HCT116 cells. (A) CDH1 KO was confirmed by western blotting. (B) Proliferation was similar between two cell types. (C) Respective cell viability after treatment with an anticancer drug, 5-FU, epidermal growth factor receptor inhibitor (erlotinib) or src kinase inhibitor (saracatinib) was similar between two cell lines. ***P<0.001 vs. WT; #P<0.05, ##P<0.01, ###P<0.001 vs. vehicle. WT, wild-type; KO, knock-out; 5-FU, 5-fluorouracil.
Fig 3: Effects of exogenous E-cadherin and DECMA-1 on WT and CDH1 KO HCT116 cells. (A) Although exogenous E-cadherin decreased viability and sizes of spheroids in both cells, a little but considerable changes was observed in CDH1 KO cells at higher dose (100 ng/ml). (B) Exogenous DECMA-1 significantly decreased viability and sizes of spheroids of both cells. Scale bar=200 µm. *P<0.05, **P<0.01, ***P<0.001 vs. vehicle; ##P<0.01 vs. WT at same concentration. WT, wild-type; KO, knock-out.
Fig 4: Effects of PD98059 on E-cadherin and spheroidogeneis of WT and CDH1 KO HCT116 cells. (A) PD98059 decreased cell viability in dose-dependent manner with a significant difference between WT and CDH1 KO HCT116 cells while ERK activation was similar. (B) PD98059 (20 µM) inhibited ERK activation in both cells, but increased E-cadherin expression. (C) PD98059 decreased sizes of spheroids in both cell lines. (D) PD98059 significantly increased the expression of β-catenin and pan-RAS without affecting the expression of EGFR. Scale bar=200 µm. *P<0.05, **P<0.01, ***P<0.001 vs. vehicle; #P<0.05, ###P<0.001 vs. WT. WT, wild-type; KO, knock-out.
Supplier Page from Abcam for Recombinant Human E Cadherin protein