The Impact of Hypoxic treatment on the Expression of Phosphoglycerate Kinase (PGK) and the Cytotoxicity of L-OddC (Troxacitabine) and dFdC (Gemcitabine)

Data Supplement

Files in this Data Supplement:

• Data Supplement - Supplemental Figure 1. Growth rate of tumor cell lines in the normoxic and hypoxic conditions (1% O2, 5% CO2, 94% N2). HepG2, Hep3B, Panc-1 and A673 cells were grown in culture medium supplemented with glucose (4.5g/L) and Hepes buffer (25mM).
• Data Supplement - Supplemental Figure 2. The time dependent uptake and incorporation of thymidine of Panc-1 cells in the normoxic and the hypoxic conditions (1% O2, 5% CO2, 94% N2). The rest of the procedures were as described in Materials and Methods.
• Data Supplement - Supplemental Figure 3. The metabolism of L-OddC and dFdC of Panc-1 cells under the normoxic and hypoxic conditions (1% O2, 5% CO2, 94% N2). Cells pre-treated under the hypoxic condition for 24h were incubated with 3HL-OddC at 500nM (2Ci/mmole) for desired time and 3HdFdC at 500nM (2Ci/mmole) for desired time. 15% TCA was used to precipitate macromolecules in the cells, the L-OddC, L-OddCMP, L-OddCDP, dFdC, dFdCMP, dFdCDP in the TCA soluble fraction were separated by HPLC (A-F). The rest of the procedures were as described in Materials and Methods.
• Data Supplement - Supplemental Figure 4. The metabolites of L-OddC (A) and dFdC (B) of Panc-1 cells under the normoxic and hypoxic conditions (1% O2, 5% CO2, 94% N2) with or without addition of DMOG 500µM. The rest of the procedures were as described in Materials and Methods.
• Data Supplement - Supplemental Figure 5. Effect of over-expression of PGK on the levels of L-OddCTP and dFdCTP and their DNA incorporation under normoxic and hypoxic conditions in RKO cells (First column). Effect of induction shHIF-1α using doxcycline on the HIF-1α and PGK expression and the level of L-OddC, dFdCTP and their DNA incorporation under normoxic and hypoxic conditions in RKO cells (Third column). Effect of the induction of shPGK using doxcycline on PGK expression and the levels of L-OddC, dFdCTP and their DNA incorporation under normoxic and hypoxic conditions RKO cells (Fourth column).
• Data Supplement - Supplemental Figure 6. Effect of over-expression of PGK on the metabolites of L-OddC under normoxic and hypoxic conditions in RKO cells (A). Effect of the induction of sh-control using doxcycline on the metabolites of L-OddC under normoxic and hypoxic conditions in RKO cells (B). Effect of the induction of shHIF-1α using doxcycline on the metabolites of L-OddC under normoxic and hypoxic conditions in RKO cells. (C). Effect of the induction of shPGK using doxcycline the metabolites of L-OddC under normoxic and hypoxic conditions in RKO cells (D).
• Data Supplement - Supplemental Figure 7. Effect of over-expression of PGK on the metabolites of dFdC under normoxic and hypoxic conditions in RKO cells (A). Effect of the induction of sh-control using doxcycline on the metabolites of dFdC under normoxic and hypoxic conditions in RKO cells (B). Effect of the induction of shHIF-1α using doxcycline on the metabolites of dFdC under normoxic and hypoxic conditions in RKO cells (C). Effect of the induction of shPGK using doxcycline the metabolites of dFdC under normoxic and hypoxic conditions in RKO cells (D).