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Semiquantification of opioid receptor mRNA expression after DMI treatment. Expression levels (at a number of thermocycles determined to fall within the linear range for both GAPDH and opioid receptor cDNA amplification) are shown as a function of time of DMI treatment. Product (5 μl) from each PCR reaction (RTs from C6 and DMI-treated C6) was taken over a range of 15-35 thermocycles and analyzed via agarose electrophoresis, followed by Southern blotting using radiolabeled probes specific for GAPDH, and the μ- or κ-opioid receptor cDNA sequence amplified. Each blot was then analyzed with the phosphorimaging program <t>ImageQuant.</t> Normalization of opioid receptor expression is given as the density of the receptor band divided by the density of the GAPDH band. A: Normalized κ expression after 0, 0.5, 1, 1.5, 2, and 20 h of DMI treatment. Shown is the comparison at 24 cycles, a point within the linear part of the amplification curve. B: Normalized μ expression after 0, 0.5, 1, 1.5, 2, and 20 h of DMI treatment. Shown is the comparison at 24 cycles, a point within the linear amplification curve. This experiment was repeated in triplicate, using different preparations of control and treated C6 cells. Data represent the average ± SEM values of three experiments.
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Semiquantification of opioid receptor mRNA expression after DMI treatment. Expression levels (at a number of thermocycles determined to fall within the linear range for both GAPDH and opioid receptor cDNA amplification) are shown as a function of time of DMI treatment. Product (5 μl) from each PCR reaction (RTs from C6 and DMI-treated C6) was taken over a range of 15-35 thermocycles and analyzed via agarose electrophoresis, followed by Southern blotting using radiolabeled probes specific for GAPDH, and the μ- or κ-opioid receptor cDNA sequence amplified. Each blot was then analyzed with the phosphorimaging program <t>ImageQuant.</t> Normalization of opioid receptor expression is given as the density of the receptor band divided by the density of the GAPDH band. A: Normalized κ expression after 0, 0.5, 1, 1.5, 2, and 20 h of DMI treatment. Shown is the comparison at 24 cycles, a point within the linear part of the amplification curve. B: Normalized μ expression after 0, 0.5, 1, 1.5, 2, and 20 h of DMI treatment. Shown is the comparison at 24 cycles, a point within the linear amplification curve. This experiment was repeated in triplicate, using different preparations of control and treated C6 cells. Data represent the average ± SEM values of three experiments.
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Semiquantification of opioid receptor mRNA expression after DMI treatment. Expression levels (at a number of thermocycles determined to fall within the linear range for both GAPDH and opioid receptor cDNA amplification) are shown as a function of time of DMI treatment. Product (5 μl) from each PCR reaction (RTs from C6 and DMI-treated C6) was taken over a range of 15-35 thermocycles and analyzed via agarose electrophoresis, followed by Southern blotting using radiolabeled probes specific for GAPDH, and the μ- or κ-opioid receptor cDNA sequence amplified. Each blot was then analyzed with the phosphorimaging program <t>ImageQuant.</t> Normalization of opioid receptor expression is given as the density of the receptor band divided by the density of the GAPDH band. A: Normalized κ expression after 0, 0.5, 1, 1.5, 2, and 20 h of DMI treatment. Shown is the comparison at 24 cycles, a point within the linear part of the amplification curve. B: Normalized μ expression after 0, 0.5, 1, 1.5, 2, and 20 h of DMI treatment. Shown is the comparison at 24 cycles, a point within the linear amplification curve. This experiment was repeated in triplicate, using different preparations of control and treated C6 cells. Data represent the average ± SEM values of three experiments.
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Semiquantification of opioid receptor mRNA expression after DMI treatment. Expression levels (at a number of thermocycles determined to fall within the linear range for both GAPDH and opioid receptor cDNA amplification) are shown as a function of time of DMI treatment. Product (5 μl) from each PCR reaction (RTs from C6 and DMI-treated C6) was taken over a range of 15-35 thermocycles and analyzed via agarose electrophoresis, followed by Southern blotting using radiolabeled probes specific for GAPDH, and the μ- or κ-opioid receptor cDNA sequence amplified. Each blot was then analyzed with the phosphorimaging program <t>ImageQuant.</t> Normalization of opioid receptor expression is given as the density of the receptor band divided by the density of the GAPDH band. A: Normalized κ expression after 0, 0.5, 1, 1.5, 2, and 20 h of DMI treatment. Shown is the comparison at 24 cycles, a point within the linear part of the amplification curve. B: Normalized μ expression after 0, 0.5, 1, 1.5, 2, and 20 h of DMI treatment. Shown is the comparison at 24 cycles, a point within the linear amplification curve. This experiment was repeated in triplicate, using different preparations of control and treated C6 cells. Data represent the average ± SEM values of three experiments.
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Semiquantification of opioid receptor mRNA expression after DMI treatment. Expression levels (at a number of thermocycles determined to fall within the linear range for both GAPDH and opioid receptor cDNA amplification) are shown as a function of time of DMI treatment. Product (5 μl) from each PCR reaction (RTs from C6 and DMI-treated C6) was taken over a range of 15-35 thermocycles and analyzed via agarose electrophoresis, followed by Southern blotting using radiolabeled probes specific for GAPDH, and the μ- or κ-opioid receptor cDNA sequence amplified. Each blot was then analyzed with the phosphorimaging program <t>ImageQuant.</t> Normalization of opioid receptor expression is given as the density of the receptor band divided by the density of the GAPDH band. A: Normalized κ expression after 0, 0.5, 1, 1.5, 2, and 20 h of DMI treatment. Shown is the comparison at 24 cycles, a point within the linear part of the amplification curve. B: Normalized μ expression after 0, 0.5, 1, 1.5, 2, and 20 h of DMI treatment. Shown is the comparison at 24 cycles, a point within the linear amplification curve. This experiment was repeated in triplicate, using different preparations of control and treated C6 cells. Data represent the average ± SEM values of three experiments.
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Semiquantification of opioid receptor mRNA expression after DMI treatment. Expression levels (at a number of thermocycles determined to fall within the linear range for both GAPDH and opioid receptor cDNA amplification) are shown as a function of time of DMI treatment. Product (5 μl) from each PCR reaction (RTs from C6 and DMI-treated C6) was taken over a range of 15-35 thermocycles and analyzed via agarose electrophoresis, followed by Southern blotting using radiolabeled probes specific for GAPDH, and the μ- or κ-opioid receptor cDNA sequence amplified. Each blot was then analyzed with the phosphorimaging program <t>ImageQuant.</t> Normalization of opioid receptor expression is given as the density of the receptor band divided by the density of the GAPDH band. A: Normalized κ expression after 0, 0.5, 1, 1.5, 2, and 20 h of DMI treatment. Shown is the comparison at 24 cycles, a point within the linear part of the amplification curve. B: Normalized μ expression after 0, 0.5, 1, 1.5, 2, and 20 h of DMI treatment. Shown is the comparison at 24 cycles, a point within the linear amplification curve. This experiment was repeated in triplicate, using different preparations of control and treated C6 cells. Data represent the average ± SEM values of three experiments.
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Semiquantification of opioid receptor mRNA expression after DMI treatment. Expression levels (at a number of thermocycles determined to fall within the linear range for both GAPDH and opioid receptor cDNA amplification) are shown as a function of time of DMI treatment. Product (5 μl) from each PCR reaction (RTs from C6 and DMI-treated C6) was taken over a range of 15-35 thermocycles and analyzed via agarose electrophoresis, followed by Southern blotting using radiolabeled probes specific for GAPDH, and the μ- or κ-opioid receptor cDNA sequence amplified. Each blot was then analyzed with the phosphorimaging program <t>ImageQuant.</t> Normalization of opioid receptor expression is given as the density of the receptor band divided by the density of the GAPDH band. A: Normalized κ expression after 0, 0.5, 1, 1.5, 2, and 20 h of DMI treatment. Shown is the comparison at 24 cycles, a point within the linear part of the amplification curve. B: Normalized μ expression after 0, 0.5, 1, 1.5, 2, and 20 h of DMI treatment. Shown is the comparison at 24 cycles, a point within the linear amplification curve. This experiment was repeated in triplicate, using different preparations of control and treated C6 cells. Data represent the average ± SEM values of three experiments.
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Semiquantification of opioid receptor mRNA expression after DMI treatment. Expression levels (at a number of thermocycles determined to fall within the linear range for both GAPDH and opioid receptor cDNA amplification) are shown as a function of time of DMI treatment. Product (5 μl) from each PCR reaction (RTs from C6 and DMI-treated C6) was taken over a range of 15-35 thermocycles and analyzed via agarose electrophoresis, followed by Southern blotting using radiolabeled probes specific for GAPDH, and the μ- or κ-opioid receptor cDNA sequence amplified. Each blot was then analyzed with the phosphorimaging program <t>ImageQuant.</t> Normalization of opioid receptor expression is given as the density of the receptor band divided by the density of the GAPDH band. A: Normalized κ expression after 0, 0.5, 1, 1.5, 2, and 20 h of DMI treatment. Shown is the comparison at 24 cycles, a point within the linear part of the amplification curve. B: Normalized μ expression after 0, 0.5, 1, 1.5, 2, and 20 h of DMI treatment. Shown is the comparison at 24 cycles, a point within the linear amplification curve. This experiment was repeated in triplicate, using different preparations of control and treated C6 cells. Data represent the average ± SEM values of three experiments.
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Semiquantification of opioid receptor mRNA expression after DMI treatment. Expression levels (at a number of thermocycles determined to fall within the linear range for both GAPDH and opioid receptor cDNA amplification) are shown as a function of time of DMI treatment. Product (5 μl) from each PCR reaction (RTs from C6 and DMI-treated C6) was taken over a range of 15-35 thermocycles and analyzed via agarose electrophoresis, followed by Southern blotting using radiolabeled probes specific for GAPDH, and the μ- or κ-opioid receptor cDNA sequence amplified. Each blot was then analyzed with the phosphorimaging program <t>ImageQuant.</t> Normalization of opioid receptor expression is given as the density of the receptor band divided by the density of the GAPDH band. A: Normalized κ expression after 0, 0.5, 1, 1.5, 2, and 20 h of DMI treatment. Shown is the comparison at 24 cycles, a point within the linear part of the amplification curve. B: Normalized μ expression after 0, 0.5, 1, 1.5, 2, and 20 h of DMI treatment. Shown is the comparison at 24 cycles, a point within the linear amplification curve. This experiment was repeated in triplicate, using different preparations of control and treated C6 cells. Data represent the average ± SEM values of three experiments.
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Semiquantification of opioid receptor mRNA expression after DMI treatment. Expression levels (at a number of thermocycles determined to fall within the linear range for both GAPDH and opioid receptor cDNA amplification) are shown as a function of time of DMI treatment. Product (5 μl) from each PCR reaction (RTs from C6 and DMI-treated C6) was taken over a range of 15-35 thermocycles and analyzed via agarose electrophoresis, followed by Southern blotting using radiolabeled probes specific for GAPDH, and the μ- or κ-opioid receptor cDNA sequence amplified. Each blot was then analyzed with the phosphorimaging program <t>ImageQuant.</t> Normalization of opioid receptor expression is given as the density of the receptor band divided by the density of the GAPDH band. A: Normalized κ expression after 0, 0.5, 1, 1.5, 2, and 20 h of DMI treatment. Shown is the comparison at 24 cycles, a point within the linear part of the amplification curve. B: Normalized μ expression after 0, 0.5, 1, 1.5, 2, and 20 h of DMI treatment. Shown is the comparison at 24 cycles, a point within the linear amplification curve. This experiment was repeated in triplicate, using different preparations of control and treated C6 cells. Data represent the average ± SEM values of three experiments.
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Semiquantification of opioid receptor mRNA expression after DMI treatment. Expression levels (at a number of thermocycles determined to fall within the linear range for both GAPDH and opioid receptor cDNA amplification) are shown as a function of time of DMI treatment. Product (5 μl) from each PCR reaction (RTs from C6 and DMI-treated C6) was taken over a range of 15-35 thermocycles and analyzed via agarose electrophoresis, followed by Southern blotting using radiolabeled probes specific for GAPDH, and the μ- or κ-opioid receptor cDNA sequence amplified. Each blot was then analyzed with the phosphorimaging program <t>ImageQuant.</t> Normalization of opioid receptor expression is given as the density of the receptor band divided by the density of the GAPDH band. A: Normalized κ expression after 0, 0.5, 1, 1.5, 2, and 20 h of DMI treatment. Shown is the comparison at 24 cycles, a point within the linear part of the amplification curve. B: Normalized μ expression after 0, 0.5, 1, 1.5, 2, and 20 h of DMI treatment. Shown is the comparison at 24 cycles, a point within the linear amplification curve. This experiment was repeated in triplicate, using different preparations of control and treated C6 cells. Data represent the average ± SEM values of three experiments.
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Semiquantification of opioid receptor mRNA expression after DMI treatment. Expression levels (at a number of thermocycles determined to fall within the linear range for both GAPDH and opioid receptor cDNA amplification) are shown as a function of time of DMI treatment. Product (5 μl) from each PCR reaction (RTs from C6 and DMI-treated C6) was taken over a range of 15-35 thermocycles and analyzed via agarose electrophoresis, followed by Southern blotting using radiolabeled probes specific for GAPDH, and the μ- or κ-opioid receptor cDNA sequence amplified. Each blot was then analyzed with the phosphorimaging program <t>ImageQuant.</t> Normalization of opioid receptor expression is given as the density of the receptor band divided by the density of the GAPDH band. A: Normalized κ expression after 0, 0.5, 1, 1.5, 2, and 20 h of DMI treatment. Shown is the comparison at 24 cycles, a point within the linear part of the amplification curve. B: Normalized μ expression after 0, 0.5, 1, 1.5, 2, and 20 h of DMI treatment. Shown is the comparison at 24 cycles, a point within the linear amplification curve. This experiment was repeated in triplicate, using different preparations of control and treated C6 cells. Data represent the average ± SEM values of three experiments.
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Image Search Results


Semiquantification of opioid receptor mRNA expression after DMI treatment. Expression levels (at a number of thermocycles determined to fall within the linear range for both GAPDH and opioid receptor cDNA amplification) are shown as a function of time of DMI treatment. Product (5 μl) from each PCR reaction (RTs from C6 and DMI-treated C6) was taken over a range of 15-35 thermocycles and analyzed via agarose electrophoresis, followed by Southern blotting using radiolabeled probes specific for GAPDH, and the μ- or κ-opioid receptor cDNA sequence amplified. Each blot was then analyzed with the phosphorimaging program ImageQuant. Normalization of opioid receptor expression is given as the density of the receptor band divided by the density of the GAPDH band. A: Normalized κ expression after 0, 0.5, 1, 1.5, 2, and 20 h of DMI treatment. Shown is the comparison at 24 cycles, a point within the linear part of the amplification curve. B: Normalized μ expression after 0, 0.5, 1, 1.5, 2, and 20 h of DMI treatment. Shown is the comparison at 24 cycles, a point within the linear amplification curve. This experiment was repeated in triplicate, using different preparations of control and treated C6 cells. Data represent the average ± SEM values of three experiments.

Journal:

Article Title: Evidence for κ - and μ -Opioid Receptor Expression in C6 Glioma Cells

doi:

Figure Lengend Snippet: Semiquantification of opioid receptor mRNA expression after DMI treatment. Expression levels (at a number of thermocycles determined to fall within the linear range for both GAPDH and opioid receptor cDNA amplification) are shown as a function of time of DMI treatment. Product (5 μl) from each PCR reaction (RTs from C6 and DMI-treated C6) was taken over a range of 15-35 thermocycles and analyzed via agarose electrophoresis, followed by Southern blotting using radiolabeled probes specific for GAPDH, and the μ- or κ-opioid receptor cDNA sequence amplified. Each blot was then analyzed with the phosphorimaging program ImageQuant. Normalization of opioid receptor expression is given as the density of the receptor band divided by the density of the GAPDH band. A: Normalized κ expression after 0, 0.5, 1, 1.5, 2, and 20 h of DMI treatment. Shown is the comparison at 24 cycles, a point within the linear part of the amplification curve. B: Normalized μ expression after 0, 0.5, 1, 1.5, 2, and 20 h of DMI treatment. Shown is the comparison at 24 cycles, a point within the linear amplification curve. This experiment was repeated in triplicate, using different preparations of control and treated C6 cells. Data represent the average ± SEM values of three experiments.

Article Snippet: Band densities were quantified with the phorphorimager analysis program ImageQuant by Molecular Dynamics.

Techniques: Expressing, Amplification, Electrophoresis, Southern Blot, Sequencing, Comparison, Control