wild type glut1  (OriGene)

Journal: Molecular medicine reports

Article Title: Combination of temozolomide and Taxol exerts a synergistic inhibitory effect on Taxol‑resistant glioma cells via inhibition of glucose metabolism.

doi: 10.3892/mmr.2015.4405

Figure Lengend Snippet: Figure 3. Taxol‑resistant cells exhibit upregulated glucose metabolism. (A) Generation of Taxol‑resistant cells from U251 glioma cells. Cells were treated with gradually increasing concentrations of Taxol in regular cell culture conditions for the selection of resistant cells. U251 Taxol‑resistant clones were pooled and analyzed following treatment with Taxol at the indicated concentrations for 48 h, and cell viability was measured. (B) Glucose uptake (Left) and lactate production (right) were measured in U251 Taxol‑sensitive and resistant cells. (C) Western blotting demonstrated that the expression levels of Glut1 and PDK1 were upregulated in Taxol‑resistant cells, as compared with the parental cells. β‑actin served as a loading control. Data are presented as the mean ± standard error of three indepen dent experiments. *P<0.05; **P<0.01. PDK1, phosphoinositide‑dependent kinase‑1; Glut1, glucose transporter 1; S, sensitive; R, resistant.

Article Snippet: A vector containing Myc-DDK-tagged wild-type Glut1 (cat. no. RC222696), was purchased from OriGene Technologies, Inc., Rockville, MD, USA).

Techniques: Cell Culture, Selection, Clone Assay, Western Blot, Expressing, Control

Journal: Molecular medicine reports

Article Title: Combination of temozolomide and Taxol exerts a synergistic inhibitory effect on Taxol‑resistant glioma cells via inhibition of glucose metabolism.

doi: 10.3892/mmr.2015.4405

Figure Lengend Snippet: Figure 2. TMZ‑resistant cells exhibit downregulated glucose metabolism. (A) Generation of TMZ‑resistant cells from U251 glioma cells. Cells were treated with gradually increasing concentrations of TMZ in regular cell culture conditions for the selection of resistant cells. U251 TMZ‑resistant clones were pooled and analyzed following treatment with TMZ at the indicated concentrations for 48 h, and cell viability was measured. (B) Glucose uptake (left) and lactate production (right) were measured in U251 TMZ‑sensitive and resistant cells. (C) Western blotting demonstrated that the expression of Glut1 and PDK1 were downregulated in TMZ‑resistant cells, as compared with the parental cells. β‑actin served as a loading control. Data are presented as the mean ± standard error of three independent experiments. *P<0.05; **P<0.01. TMZ, temozolomide; PDK1, phosphoinositide‑dependent kinase‑1; Glut1, glucose transporter 1; S, sensitive; R, resistant.

Article Snippet: A vector containing Myc-DDK-tagged wild-type Glut1 (cat. no. RC222696), was purchased from OriGene Technologies, Inc., Rockville, MD, USA).

Techniques: Cell Culture, Selection, Clone Assay, Western Blot, Expressing, Control

Journal: Molecular medicine reports

Article Title: Combination of temozolomide and Taxol exerts a synergistic inhibitory effect on Taxol‑resistant glioma cells via inhibition of glucose metabolism.

doi: 10.3892/mmr.2015.4405

Figure Lengend Snippet: Figure 5. Overexpression of Glut1 renders Taxol‑resistant cells unsusceptible to Taxol and TMZ combined treatment. (A) Treatment with a combination of Taxol and TMZ inhibited glucose uptake (left) and lactate production (right). Taxol‑resistant U251 glioma cells were treated with Taxol alone at 1 µM; TMZ alone at 5 µM, or Taxol + TMZ for 48 h, following which glucose uptake (left) and lactate production (right) were measured. (B) Western blotting demonstrated that the expression levels of Glut1 in U251 Taxol‑resistant cells transiently transfected with Glut1 were higher, as compared with cells transfected with a vector control (left). U251 Taxol‑resistant cells were transiently transfected with vector control or wild‑type Glut1 for 48 h, and were plated into 48‑well plates for the indicated treatments for 48 h, followed by measurements of cell viability. β‑actin served as a loading control. Data are presented as the mean ± standard error of three independent experiments. *P<0.05. TMZ, temozolomide; Ctrl, control; R, resistant; Glut1, glucose transporter 1; V, vector.

Article Snippet: A vector containing Myc-DDK-tagged wild-type Glut1 (cat. no. RC222696), was purchased from OriGene Technologies, Inc., Rockville, MD, USA).

Techniques: Over Expression, Western Blot, Expressing, Transfection, Plasmid Preparation, Control

Journal: Molecular medicine reports

Article Title: Combination of temozolomide and Taxol exerts a synergistic inhibitory effect on Taxol‑resistant glioma cells via inhibition of glucose metabolism.

doi: 10.3892/mmr.2015.4405

Figure Lengend Snippet: Figure 3. Taxol‑resistant cells exhibit upregulated glucose metabolism. (A) Generation of Taxol‑resistant cells from U251 glioma cells. Cells were treated with gradually increasing concentrations of Taxol in regular cell culture conditions for the selection of resistant cells. U251 Taxol‑resistant clones were pooled and analyzed following treatment with Taxol at the indicated concentrations for 48 h, and cell viability was measured. (B) Glucose uptake (Left) and lactate production (right) were measured in U251 Taxol‑sensitive and resistant cells. (C) Western blotting demonstrated that the expression levels of Glut1 and PDK1 were upregulated in Taxol‑resistant cells, as compared with the parental cells. β‑actin served as a loading control. Data are presented as the mean ± standard error of three indepen dent experiments. *P<0.05; **P<0.01. PDK1, phosphoinositide‑dependent kinase‑1; Glut1, glucose transporter 1; S, sensitive; R, resistant.

Article Snippet: A vector containing wild-type Glut1 was purchased from OriGene Technologies, Inc. (cat. no. RC222696).

Techniques: Cell Culture, Selection, Clone Assay, Western Blot, Expressing, Control

Journal: Molecular medicine reports

Article Title: Combination of temozolomide and Taxol exerts a synergistic inhibitory effect on Taxol‑resistant glioma cells via inhibition of glucose metabolism.

doi: 10.3892/mmr.2015.4405

Figure Lengend Snippet: Figure 2. TMZ‑resistant cells exhibit downregulated glucose metabolism. (A) Generation of TMZ‑resistant cells from U251 glioma cells. Cells were treated with gradually increasing concentrations of TMZ in regular cell culture conditions for the selection of resistant cells. U251 TMZ‑resistant clones were pooled and analyzed following treatment with TMZ at the indicated concentrations for 48 h, and cell viability was measured. (B) Glucose uptake (left) and lactate production (right) were measured in U251 TMZ‑sensitive and resistant cells. (C) Western blotting demonstrated that the expression of Glut1 and PDK1 were downregulated in TMZ‑resistant cells, as compared with the parental cells. β‑actin served as a loading control. Data are presented as the mean ± standard error of three independent experiments. *P<0.05; **P<0.01. TMZ, temozolomide; PDK1, phosphoinositide‑dependent kinase‑1; Glut1, glucose transporter 1; S, sensitive; R, resistant.

Article Snippet: A vector containing wild-type Glut1 was purchased from OriGene Technologies, Inc. (cat. no. RC222696).

Techniques: Cell Culture, Selection, Clone Assay, Western Blot, Expressing, Control

Journal: Molecular medicine reports

Article Title: Combination of temozolomide and Taxol exerts a synergistic inhibitory effect on Taxol‑resistant glioma cells via inhibition of glucose metabolism.

doi: 10.3892/mmr.2015.4405

Figure Lengend Snippet: Figure 5. Overexpression of Glut1 renders Taxol‑resistant cells unsusceptible to Taxol and TMZ combined treatment. (A) Treatment with a combination of Taxol and TMZ inhibited glucose uptake (left) and lactate production (right). Taxol‑resistant U251 glioma cells were treated with Taxol alone at 1 µM; TMZ alone at 5 µM, or Taxol + TMZ for 48 h, following which glucose uptake (left) and lactate production (right) were measured. (B) Western blotting demonstrated that the expression levels of Glut1 in U251 Taxol‑resistant cells transiently transfected with Glut1 were higher, as compared with cells transfected with a vector control (left). U251 Taxol‑resistant cells were transiently transfected with vector control or wild‑type Glut1 for 48 h, and were plated into 48‑well plates for the indicated treatments for 48 h, followed by measurements of cell viability. β‑actin served as a loading control. Data are presented as the mean ± standard error of three independent experiments. *P<0.05. TMZ, temozolomide; Ctrl, control; R, resistant; Glut1, glucose transporter 1; V, vector.

Article Snippet: A vector containing wild-type Glut1 was purchased from OriGene Technologies, Inc. (cat. no. RC222696).

Techniques: Over Expression, Western Blot, Expressing, Transfection, Plasmid Preparation, Control