Introduction of the iHAC to HFL-1 by MV-MMCT using Haals-αTfR. (A) Representative bright-field and fluorescence images of an EGFP(+) colony. Three dedifferentiated clones (h-A9-3, h-A2 and h-A8) exhibited doom-like morphology, whereas a clone h-D20 still retained fibroblast-like morphology. (B) Expression of exogenous reprogramming factors detected by RT-PCR in EGFP(+)/dedifferentiated clones. NAT1 was used as an internal control. (C) FISH analysis of EGFP(+) clone. Digoxigenin-labeled alphoid satellite marker (red) was used to detect the HAC backbone and endogenous chromosomes 13 and 21. Biotin-labeled pPAC-2CAG-O2hP53sh (green) was used to detect the reprogramming cassette in the iHAC. Chromosomal DNA was counterstained with DAPI. White arrows indicate iHAC vector and the insets show enlarged images of the iHACs. Arrowheads indicate Chr.13 or 21.