Figure 3. YAP1 knockdown in CD133⁺CD44⁺ or CD133^-CD44^- CRC cells affected their proliferation, CD133⁺CD44⁺ percentages, relative gene expression.

(A) Western blots of YAP1 in CD133⁺CD44⁺ or CD133^-CD44^- CRC cells transiently transfected with YAP1 siRNA1 (siRNA-YAP1-1), siRNA2 (siRNA-YAP1-2) or siRNA3 (siRNA-YAP1-3); β-actin was used as a loading control. (B and C) The proliferation rates of CD133⁺CD44⁺ and CD133^-CD44^- HT-29 (B) or Caco-2 (C) cells transfected with siRNA-YAP1-1 from days 1 through 4 after seeding (^**: P<0.01). (D and E) The CD133⁺CD44⁺ percentages determined by flow cytometry in YAP1-interfered CD133⁺CD44⁺ or CD133^-CD44^- CRC cells. (F) The protein expression levels of Ascl2, KLF5 and ‘stemness’-associated genes in the whole cell lysates of CD133⁺CD44⁺ CRC cells transfected with siRNA-YAP1-1; β-actin was used as a loading control. YAP1 nuclear accumulation was inhibited in siRNA-YAP1-1-transfected CD133⁺CD44⁺ CRC cells; Lamin B1 was used as an internal control for the nuclear fraction.