Journal: Cell Chemical Biology

Article Title: Targeting non-canonical pathways as a strategy to modulate the sodium iodide symporter

doi: 10.1016/j.chembiol.2021.07.016

Figure Lengend Snippet:

Article Snippet: Phenformin HCL , Stratech Scientific , Cat# B1373.

Techniques: Virus, Subcloning, Recombinant, Saline, Reverse Transcription, Plasmid Preparation, Gene Expression, Negative Control, Positive Control, Software, Drug discovery, High Content Screening, Real-time Polymerase Chain Reaction

Journal: International Journal of Molecular Sciences

Article Title: The Oligostilbene Gnetin H Is a Novel Glycolysis Inhibitor That Regulates Thioredoxin Interacting Protein Expression and Synergizes with OXPHOS Inhibitor in Cancer Cells

doi: 10.3390/ijms24097741

Figure Lengend Snippet: Effects on T98G cells of combined treatment with glycolysis inhibitors and the mitochondrial complex-I inhibitor phenformin. ( A ) T98G cells treated for 48 h with either 4 µM GH or other known glycolysis inhibitors, such as 25 mM dichloroacetate (DCA), 25 mM oxamate, 25 mM 2-deoxy- d -glucose (2DG), 25 µM 3- bromopyruvate (3BrPA), alone or in combination with 100 µM of the mitochondrial complex-I inhibitor phenformin. Data is representative of three biological repeats (±SEM) 2-way ANOVA/Tukey’s multiple comparisons test; * p ≤ 0.05, *** p ≤ 0.001, **** p ≤ 0.0001 ( B ) Assessment of apoptosis (early/late apoptosis) and necrosis following treatment with 4 µM GH alone or in combination with 100 µM phenformin for 48 h by annexin-V/propidium iodide (PI) staining. ( C ) Analysis of T98G viability independent from the effect on cell metabolism using a protease activity-based cell viability assay. T98G cells were treated for 3 days with 8 µM GH, 100 µM phenformin, or the combination of the two was assessed at 24 h. Data is representative of three biological repeats, analyzed by 2-WAY ANOVA/Dunnett’s multiple comparisons test; CTRL VH vs. GH/time point, * p ≤ 0.05,** p ≤ 0.01, *** p ≤ 0.001; CTRL VH vs. Phen/time point, ### p ≤ 0.001; CTRL VH vs. GH + Phen/time point, ◊◊ p ≤ 0.01, ◊◊◊ p ≤ 0.001.

Article Snippet: Dichloroacetate (DCA), oxamate, 2-deoxy- d -glucose (2DG), 3-bromopyruvate (3BrPA), and phenformin (Phen) were purchased from Cayman Chemicals (Ann Arbor, MI, USA).

Techniques: Staining, Activity Assay, Viability Assay

Journal: International Journal of Molecular Sciences

Article Title: The Oligostilbene Gnetin H Is a Novel Glycolysis Inhibitor That Regulates Thioredoxin Interacting Protein Expression and Synergizes with OXPHOS Inhibitor in Cancer Cells

doi: 10.3390/ijms24097741

Figure Lengend Snippet: The treatment with GH impairs the glycolytic activity of T98G cells. The glycolytic activity was measured by Agilent SeaHorse XFe96 Analyzer after 20 min of treatment with either phenformin (100 µM), gnetin H (10 µM), or the combination injected into the cell media following basal ECAR measurements. (*) indicates the time of treatment with phen, GH, or the combination. ECAR data are shown as mpH/min normalized to proteins and represent the mean ± SD of n = 5 independent measurements under normal cell culture conditions. Glycolytic profiles were obtained using the Agilent SeaHorse Glycolysis Stress Test. The ECAR was measured under glucose starvation and after the sequential addition of glucose (basal glycolysis), oligomycin (maximal glycolysis or glycolytic reserve), and 2-DG (glycolysis specificity).

Article Snippet: Dichloroacetate (DCA), oxamate, 2-deoxy- d -glucose (2DG), 3-bromopyruvate (3BrPA), and phenformin (Phen) were purchased from Cayman Chemicals (Ann Arbor, MI, USA).

Techniques: Activity Assay, Injection, Cell Culture

Journal: International Journal of Molecular Sciences

Article Title: The Oligostilbene Gnetin H Is a Novel Glycolysis Inhibitor That Regulates Thioredoxin Interacting Protein Expression and Synergizes with OXPHOS Inhibitor in Cancer Cells

doi: 10.3390/ijms24097741

Figure Lengend Snippet: RNA-Seq analysis of T98G cells treated with 2DG, Phen, 2DG + Phen, GH, or GH + Phen. ( A ) The experimental variation between RNA-Seq experiments performed on 3 biological replicate samples was analyzed with a PCA plot. ( B ) Venn diagrams comparing the number of up- or downregulated genes that significantly responded (≥2-fold induction, p -value ≤ 0.05) to the different treatments. ( C ) Hierarchical cluster analysis was performed on 566 genes that displayed a ≥2-fold induction ( p -value ≤ 0.05) response to at least one of the three treatments compared to the solvent control. Blue and yellow colors represent up- and downregulated genes, respectively. The genes listed responded to GH + Phen treatment, but not to 2DG or 2DG + Phen treatment.

Article Snippet: Dichloroacetate (DCA), oxamate, 2-deoxy- d -glucose (2DG), 3-bromopyruvate (3BrPA), and phenformin (Phen) were purchased from Cayman Chemicals (Ann Arbor, MI, USA).

Techniques: RNA Sequencing, Solvent, Control