Figure 3. MEK1 knockdown suppresses liver CSC self-renewal and tumorgenetic capacity.

(A) Western blot analysis MEK1 and the substrate ERK1/2 expression in Huh7- and PLC/PRF/5-Nanog^Pos cells which depleted MEK1 with two individual lentiviruses for 48 hours. (B) Effect of MEK1 knockdown on cellular growth rates of Huh7- and PLC/PRF/5- Nanog^Pos cells. CCK8 assay was performed after transfection with indicated times. Cell lysates were obtained from cells transiently transfected with either MEK1 shRNA or negative control shRNA. (C) Huh7- and PLC/PRF/5- Nanog^Pos cells which transfected with MEK1 shRNA or negative control shRNA cultured under non-adhesive culture system for 7 days. (D) Huh7- and PLC/PRF/5-Nanog^Pos cells were transduced with lentiviruses expressing the indicated shRNA. Cells were grown for 14 days and stained with crystal violet. (E) Western blot analysis of stemness-related proteins in MEK1-depleted cells, relative to control.