LIP induced transformation of primary BM cells in vitro in the same way as Evi1; LAP* and LAP did not. (A) Total cell lysates of PLAT-E cells transfected with pGCDNsam-LAP*, LAP, or LIP-Kusabira Orange were immunoblotted with the anti-C/EBPβ antibody (Santa Cruz, C-19). (B) Third colonies of the primary BM cells transduced with Evi1, LAP*, LAP, LIP, and mock (top panels). The Evi1-colonies were GFP positive, and the LIP-colonies were Kusabira Orange–positive (bottom panels). (C) Numbers of CFUs per cell plated in each generation. Mean from 2 independent experiments are depicted with standard deviation, each in duplicate. (D) Growth curves of the Evi1-expressing cells and the LIP-expressing cells in liquid culture (medium conditions: RPMI containing 10% fetal calf serum and 5 ng/mL of IL-3). (E-F) The liquid culture cells transformed by LIP (E) or Evi1 (F) were stained with Wright-Giemsa. Microscope, BH-2, Olympus; camera module, DP20, Olympus; objective lens, NC SPlan, Olympus; magnification, ×1000. (G) Flow cytometric analysis of the Evi1-liquid culture cells and the LIP-liquid culture cells.