Figure 2.

ECE1c^K6R expression induces stemness and spheroidogenesis. (A) DLD‐1 cells expressing either Flag‐tagged ECE1c^WT or ECE1c^K6R proteins were grown under normal conditions for 48 h, and the mRNA levels of stemness genes were quantified using RT‐qPCR (n = 3). (B) Cells grown as in A were evaluated for CD133⁺/CD44⁺ population using flow cytometry and flowjo V.10 software (n = 3). (C) As in A, cells were grown for 7 days under spheroidogenic conditions. Representative images are shown (scale bar = 150 µm). (D) Number of spheroids formed in C was determined from a triplicate analysis. (E) Sizes of spheroids formed in C were quantified from a triplicate analysis. Data represent average ± SEM (n = 3). ANOVA and Tukey tests were used, except for B: t‐test. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001.