Supplementary Table 1 - Primers used
Target sequences
Primer sequence（5’— 3’）
pTet on-hTERT a
CGTCAGATCGCCTGGAGAAT
Size(bp)
870
ACAGAAACCACGGTCACTCG
hTERT b
CCGTACATGCGACAGTTC
200
GCAGAGCAGCGTGGAGAG
GAPDH c
TGATGACATCAAGAAGGTGGTGAAG
275
TCCTTGGAGGCCATGTAGGCCAT
a
Primers are used for genomic PCR
b, c
Primers are used for RT-PCR
For each primer pair used, the sequence of the forward (top) and reverse (bottom) primers are shown.
The right column indicates the size of the expected PCR products.
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Supplementary Figure legends
Supplementary Fig. 1 The structure of the plasmid pTet-on EGFP
Supplementary Fig. 2 Proliferative capacity of the hTERT transgenic cells. (a) Growth curves of the
clones, h1, h3 and h4. The cells were cultured in medium containing doxycycline (Dox) (h(+) or without
Dox (h(-)), and their population doubling (PD) levels were evaluated at every passage untill h(-) group
cells died or almost died. h(+) cells maintained a similar growth rate to early passages of cells, while h(-)
cells showed a slower growth rate and gradually stopped proliferation. (b) SA–β-galactosidase (SA–βGal) staining of the cells. The cells were stained for SA–β-Gal at late passages. There were no positive
signals of dark blue substances for h(+) cells after more than 80 PDs (left), whereas dark blue substances
were detected in h(-) cells by 50 PDs (right). Scale bar, 50 μm
Supplementary Fig. 3 The reversal of immortalized cells. (a) Growth curves of cell lines, h1, h3 and h4.
h(+) cells maintained their vigorous growth after more than 300 PDs, while h(+-) cells showed a slower
growth rate and stopped dividing by less than 270 PDs. (d) SA–β-galactosidase (SA–β-Gal) staining of
the cells. There were no positive signals of dark blue substances for h(+) cells (left), whereas dark blue
substances were detected in h(+-) cells by 250 PDs (right). Scale bar, 50 μm
2
Supplementary Fig. 1 The structure of the plasmid pTet-on EGFP
3
Supplementary Fig. 2 Proliferative capacity of the hTERT transgenic cells
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Supplementary Fig. 3 The reversal of immortalized cells
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