Fig. 5.

Altered RT of TP63 is an early marker of progeroid diseases. (A) Reprogramming of primary cells to iPSCs and redifferentiation back to fibroblasts as a model of progeroid disease progression. (B) Representative immunofluorescence images of normal and HGPS cells confirm that reprogrammed iPSCs revert to the HGPS nuclear abnormalities which are spontaneously reestablished upon differentiation. (Magnification: 60×.) (C) Genome-wide RT correlation of primary cells, iPSCs, and redifferentiated cells from normal, HGPS, and RTS donors. (D) Clustering analysis of fibroblasts and redifferentiated cells from iPSCs. Only regions from the E-progeria RT signature are shown. (E and F) Altered TP63 RT is reestablished in redifferentiated HGPS and RTS cells. (G) TP63 isoforms expression by qRT-PCR in primary cells, iPSCs, and redifferentiated cells. Relative expression was normalized against the HPRT1 gene. Significant differences are shown (*P < 0.05). (H) TP63 changes to early replication during the first stages of mesoderm differentiation and switches back to late replication in later stages and is highly induced only when the gene is replicated early during S-phase. Re-diff, redifferentiated.