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. 2013 Jun 10;123(7):2832–2849. doi: 10.1172/JCI65859

Figure 5. Validation of Yap1 and Igf2bp3 as TIC oncogenes.

Figure 5

(A) The number of colonies formed by TICs in soft agar is moderately reduced by lentiviral transduction of shRNA against Yap1 or Igf2bp3 but largely abrogated when knocking down both. Stat3 silencing has no effects. *P < 0.05, **P < 0.01. (B) Self-renewal ability, as assessed by spheroid formation assay, is reduced in sh-Yap1/sh-Igf2bp3–transduced TICs. Serial spheroid-forming capacity is also reduced by dual transduction of sh-Yap1 and sh-Igf2bp3. *P < 0.05. (C) Immunoblot analysis demonstrates induced expression of IGF2BP3 and YAP1 in TICs and effective knockdown by respective shRNA. (D) Transduction of Ifg2bp3 shRNA, but not that of Yap1 shRNA, modestly reduces subcutaneous tumor growth by the TICs from the Ns5a Tg mice in NOG mice, while transduction of both shRNA clearly abrogates the growth at the later 3 time points. Representative photos of NOG mice carrying TIC-derived tumors are shown to depict an appreciable reduction of tumor size by Igf2bp3 and Yap1 silencing. *P < 0.05, **P < 0.01 versus scrambled shRNA group.