FIGURE 9.

YAP rescued effects of CD166 depletion. A, YAP rescued CD166 knockdown-induced down-regulation of Bcl-2. Shown are Western blots of Bcl-2 in Bel-7402 cells with CD166 knocked down (infected with CD166-sh1) with or without simultaneous overexpression of YAP (transfected with YAP-FLAG-expressing plasmids). B, knockdown of CD166 (infected with CD166-sh1) induced apoptosis, which can be rescued by overexpression of YAP (infected with YAP-expressing lentivirus), as measured by caspase-3/7 activity. C, relative cell proliferation measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide in cells expressing shRNA against CD166 (CD166-sh1) with or without ectopic expression of YAP (infected with YAP-expressing lentivirus). D, ectopic expression of YAP rescued inhibitory tumor growth by silencing of CD166 in vivo. Tumor volumes were measured for 24 days after subcutaneous injection of different cells as indicated. n = 5 per group. p < 0.05 or 0.01 indicates statistic significance. E, representative IHC images of control and xenograft tissues with CD166 knocked down from D stained using anti-CD133 and anti-Ki67 antibodies. Scale bar, 200 μm. F, Western blots of stem cell markers as indicated in control cells (infected with GFP-shRNA) and Bel-7402 cells with CD166 knocked down (infected with CD166-sh1). Error bars, S.D.