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www.impactjournals.com/oncotarget/ Oncotarget, Supplementary Materials 2015
SUPPLEMENTARY FIGURES AND TABLE
Supplementary Figure S1: Short-term chemotherapy has minimal effects on tumor cells viability. Representative ow
cytometry dot plots of BC cells upon short-term single exposure to chemotherapy. Cells were treated with 10 μM CIS or 10 μM MTX for
24 hours and then subjected to ow cytometry analysis. R1 represents the percentage of dead cells and R2 the percentage of viable cells of
the whole-cell population.
www.impactjournals.com/oncotarget/ Oncotarget, Supplementary Materials 2015
Supplementary Figure S2: Bladder cancer induced by orthotopic inoculation of adherent HT-1376 cells. A. Representative
BLI image of a mouse that was inoculated intravesically with 4×106 adherent HT-1376 cells at 5 weeks. B. Macroscopic examination of
a papillary tumor protruding into the bladder lumen on day 40. C. Representative H&E–stained (×200) and high mitotic count (×400).
D. Immunohistochemical staining for Ki67, CD44, CD47, SOX2 and ALDH2 in serial sections of normal urothelium (upper row) and in
the orthotopic tumor (lower row). Original magnication: ×400.
www.impactjournals.com/oncotarget/ Oncotarget, Supplementary Materials 2015
Supplementary Table S1: Primer sequences used in real-time RT-qPCR analysis
Gene Primer Primer Sequence/ENSEMBL Gene ID
ALDH1A1 Forward 5′-TGGCTTATCAGCAGGAGTGT-3′
ALDH1A1 Reverse 5′-GCAATTCACCCACACTGTTC-3′
ALDH2 Forward 5′-CGAGGTCTTCTGCAACCAG-3′
ALDH2 Reverse 5′-GCCTTGTCCACATCTTCCTT-3′
ALDH7A1 Forward 5′-CAACGAGCCAATAGCAAGAG-3′
ALDH7A1 Reverse 5′-GCATCGCCAATCTGTCTTAC-3′
SOX2 Forward 5′-CATGCACCGCTACGACG-3′
SOX2 Reverse 5′-CGGACTTGACCACCGAAC-3′
NANOG Forward 5′-GATGCCTCACACGGAGACTG-3′
NANOG Reverse 5′-GCAGAAGTGGGTTGTTTGCC-3′
POU5F1 Forward 5′-TCTGCATCCCTTGGATGTGC-3′
POU5F1 Reverse 5′-GTGTGGCCCCAAGGAATAGT-3′
ABCB1 Forward 5′-GAGGAAGACATCACCAGGTATGC-3′
ABCB1 Reverse 5′-GCCAGGCACCAAAATGAAACC-3′
ABCG2 Forward 5′-TCAGCGGATACTACAGAGTGTCATC-3′
ABCG2 Reverse 5′-ATCTGCCTTTGGCTTCAATCCTAA-3′
HPRT-1 Forward 5′-TGACACTGGCAAAACAATG-3′
HPRT-1 Reverse 5′-GGCTTATATCCAACACTTCG-3′
18S Forward 5′-GAAGATATGCTCATGTGGTGTTG-3′
18S Reverse 5′-CTTGTACTGGCGTGGATTCTG-3′
GAPDH Forward 5′-ACAGTCAGCCGCATCTTC-3′
GAPDH Reverse 5′-GCCCAATACGACCAAATCC-3′
CD44 ENST00000278385
CD47 ENST00000398257
KRT14 ENST00000167586
All Cycle threshold (Ct) values of the analyzed genes were normalized to the housekeeping genes, i.e., HRPT-1, 18S and
GAPDH.