Journal: PLoS ONE

Article Title: PKM2 Subcellular Localization Is Involved in Oxaliplatin Resistance Acquisition in HT29 Human Colorectal Cancer Cell Lines

doi: 10.1371/journal.pone.0123830

Figure Lengend Snippet: Dose-response curves for HT29, SW480 and HCT116 cell lines after PKM2 gene silencing and OXA treatment at 0–140 μM and 0–32 μM for 24 hours. Curves represent the average values from at least three independent experiments. Cell proliferation was measured by MTT assay. Vertical bars in the graphics represent ± SD. Insets show PKM2 inmunoblotting after siRNA-directed inhibition. Specific IC50 values for oxaliplatin in all conditions are displayed in the table. IC50 values for Mock conditions (without transfection) were very similar to those of siNTC and are not shown. *p-values are result of comparison to the siNTC condition.

Article Snippet: Cell lines were grown as monolayers in DMEM (HT29, SW480 and HTOXAR3; Invitrogen, Life Technologies) or RPMI 1640 (HCT116 and HCT116 p53 null; Invitrogen, Life Technologies) supplemented with 10% heat-inactivated FCS (Reactiva), 400 units/ml of penicillin, 40 μg/ml of gentamycin, and 2 mM of L-glutamine (Sigma) and cultured at 37°C in a humidified atmosphere of 5% CO 2 .

Techniques: MTT Assay, Inhibition, Transfection

Journal: PLoS ONE

Article Title: PKM2 Subcellular Localization Is Involved in Oxaliplatin Resistance Acquisition in HT29 Human Colorectal Cancer Cell Lines

doi: 10.1371/journal.pone.0123830

Figure Lengend Snippet: Immunoflourescence staining of PKM2 (red) demonstrates nuclear accumulation in HCT116 and HT29 cells after treatment with OXA in a time and dose-dependent manner but not in resistant HTOXAR3 cells. Nuclei were stained in blue. NT: Non-treated cells. Objective lens: 40x immersion oil. Scale bar: 10 μm.

Article Snippet: Cell lines were grown as monolayers in DMEM (HT29, SW480 and HTOXAR3; Invitrogen, Life Technologies) or RPMI 1640 (HCT116 and HCT116 p53 null; Invitrogen, Life Technologies) supplemented with 10% heat-inactivated FCS (Reactiva), 400 units/ml of penicillin, 40 μg/ml of gentamycin, and 2 mM of L-glutamine (Sigma) and cultured at 37°C in a humidified atmosphere of 5% CO 2 .

Techniques: Staining

Journal: PLoS ONE

Article Title: PKM2 Subcellular Localization Is Involved in Oxaliplatin Resistance Acquisition in HT29 Human Colorectal Cancer Cell Lines

doi: 10.1371/journal.pone.0123830

Figure Lengend Snippet: A. 3-D plot showing fold changes in expression patterns after treatment with 10 μM OXA in HT29, siPKM2-HT29 and HTOXAR3 cells. B. Heat map showing up- and down-regulated genes after OXA treatment according to three different cell death pathways.

Article Snippet: Cell lines were grown as monolayers in DMEM (HT29, SW480 and HTOXAR3; Invitrogen, Life Technologies) or RPMI 1640 (HCT116 and HCT116 p53 null; Invitrogen, Life Technologies) supplemented with 10% heat-inactivated FCS (Reactiva), 400 units/ml of penicillin, 40 μg/ml of gentamycin, and 2 mM of L-glutamine (Sigma) and cultured at 37°C in a humidified atmosphere of 5% CO 2 .

Techniques: Expressing

Journal: PLoS ONE

Article Title: PKM2 Subcellular Localization Is Involved in Oxaliplatin Resistance Acquisition in HT29 Human Colorectal Cancer Cell Lines

doi: 10.1371/journal.pone.0123830

Figure Lengend Snippet: Most relevant changes in cell death genes expression patterns after treatment with OXA and/or PKM2 gene silencing.

Article Snippet: Cell lines were grown as monolayers in DMEM (HT29, SW480 and HTOXAR3; Invitrogen, Life Technologies) or RPMI 1640 (HCT116 and HCT116 p53 null; Invitrogen, Life Technologies) supplemented with 10% heat-inactivated FCS (Reactiva), 400 units/ml of penicillin, 40 μg/ml of gentamycin, and 2 mM of L-glutamine (Sigma) and cultured at 37°C in a humidified atmosphere of 5% CO 2 .

Techniques: Expressing

Journal: PLoS ONE

Article Title: PKM2 Subcellular Localization Is Involved in Oxaliplatin Resistance Acquisition in HT29 Human Colorectal Cancer Cell Lines

doi: 10.1371/journal.pone.0123830

Figure Lengend Snippet: Changes in BMF gene expression between OXA treated (T) and non-treated (NT) HT29, siPKM2- HT29 and HTOXAR3 cells. Vertical bars in the graphics represent means obtained from at least 3 independent experiments ± SD. B. Percentage of dead cells after treatment with oxaliplatin and/or BMF gene knockdown. Bars represent means obtained from at least 3 independent experiments ± SD. The Little graph shows percentage (mean ± SD) of BMF expression inhibition after siRNA transfections. * P- value < 0.05. ** P-value < 0.01; *** P-value < 0.001

Article Snippet: Cell lines were grown as monolayers in DMEM (HT29, SW480 and HTOXAR3; Invitrogen, Life Technologies) or RPMI 1640 (HCT116 and HCT116 p53 null; Invitrogen, Life Technologies) supplemented with 10% heat-inactivated FCS (Reactiva), 400 units/ml of penicillin, 40 μg/ml of gentamycin, and 2 mM of L-glutamine (Sigma) and cultured at 37°C in a humidified atmosphere of 5% CO 2 .

Techniques: Expressing, Inhibition, Transfection