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Results: 7

1.
Figure 3

Figure 3. From: Up-regulation of the mitochondrial malate dehydrogenase by oxidative stress is mediated by miR-743a.

H2O2 reduced the miR-743a expression in HT22 cells. (A) Alignment of miR-743a with mdh2 3′ UTR. (B) H2O2 (20 μM for 8 hr) reduced the miR-743a expression. miR-743a expression was measured by real-time PCR using sno-202 as an internal control. Values are the means ± SEM of relative changes over controls from two independent experiments in triplicate after normalization to sno-202. Values with different letters vary statistically from each other (P<0.05) as determined by the unpaired two tailed Student’s T-test.

Qingli Shi, et al. J Neurochem. ;118(3):440-448.
2.
Figure 5

Figure 5. From: Up-regulation of the mitochondrial malate dehydrogenase by oxidative stress is mediated by miR-743a.

Anti-miR-743a increased luciferase activity of a dual luciferase reporter containing mdh2 3′ UTR. Co-transfection of this reporter (Luc+mdh) with an anti-miR-743a increased luciferase activity compared to the control (cells co-transfected with negative control for anti-miRNA and Luc-mdh). Co-transfection of the luciferase reporter without mdh 3′UTR (Luc-mdh) with the anti-miR-743a did not alter the luciferase activity. Values are the means ± SEM of percent changes over controls after normalization to the Renilla luciferase activity from three independent experiments in triplicate. Values with different letters vary significantly from each other (P<0.05) as determined by a one-way ANOVA followed by a Student-Newman-Keul’s test.

Qingli Shi, et al. J Neurochem. ;118(3):440-448.
3.
Figure 7

Figure 7. From: Up-regulation of the mitochondrial malate dehydrogenase by oxidative stress is mediated by miR-743a.

Endogenous mRNA (A) and activity (B) levels of MDH were up-regulated by an anti-miR-743a. Endogenous mRNA level of ogdh was not altered by anti-miR-743a (C). HT22 cells were transfected with an anti-miR-743a or a negative control, mRNA levels of mdh2 and ogdh, as well as MDH activity were measured. Values are the means ± SEM from three independent experiments in triplicate. Values with different letters vary statistically from each other (P<0.05) as determined by the unpaired two tailed Student’s T-test.

Qingli Shi, et al. J Neurochem. ;118(3):440-448.
4.
Figure 1

Figure 1. From: Up-regulation of the mitochondrial malate dehydrogenase by oxidative stress is mediated by miR-743a.

Hydrogen peroxide (H2O2) increased malate dehydrogenase (MDH) activity (A) and mRNA level (B) in HT22 cells. Cells were treated with 20 μM H2O2 for 8 hr. Values are means ± SEM from at least two independent experiments in triplicate. Values with different letters differ significantly (P < 0.05) from each other as determined by the unpaired two tailed Student’s T-test.

Qingli Shi, et al. J Neurochem. ;118(3):440-448.
5.
Figure 6

Figure 6. From: Up-regulation of the mitochondrial malate dehydrogenase by oxidative stress is mediated by miR-743a.

Endogenous mRNA (A) and activity (B) levels of MDH were diminished by miR-743a. Endogenous mRNA level of ogdh was not altered by miR-743a (C). HT22 cells were transfected with miR-743a or a negative control miRNA. mRNA levels of mdh2 and ogdh, as well as MDH activity were measured. Values are the means ± SEM from three independent experiments in triplicate. Values with different letters vary statistically from each other (P<0.05) as determined by the unpaired two tailed Student’s T-test.

Qingli Shi, et al. J Neurochem. ;118(3):440-448.
6.
Figure 2

Figure 2. From: Up-regulation of the mitochondrial malate dehydrogenase by oxidative stress is mediated by miR-743a.

H2O2-induced activation of mdh2 is likely post-transcriptional. HT22 cells were treated with H2O2 in the presence or absence of actinomycin D (Act D) and the mRNA levels of mdh2 (A) and sdha (B) were measured by real-time PCR using β-microglobulin (β2m) as an internal control. Values are mean ± SEM from at least two independent experiments in triplicate. Values with different letters differ significantly (P < 0.05) from each other as determined by a one-way ANOVA followed by a Student-Newman-Keul’s test.

Qingli Shi, et al. J Neurochem. ;118(3):440-448.
7.
Figure 4

Figure 4. From: Up-regulation of the mitochondrial malate dehydrogenase by oxidative stress is mediated by miR-743a.

miR-743a inhibited luciferase activity of a dual luciferase reporter containing mdh2 3′ UTR. The firefly luciferase was fused to the entire mdh2 mRNA 3′ UTR. Co-transfection of this reporter (Luc+mdh) with miR-743a reduced luciferase activity compared to the control (cells co-transfected with negative control miRNA and Luc-mdh). Co-transfection of the luciferase reporter without mdh 3′UTR (Luc-mdh) with miR-743a did not alter the luciferase activity. Values are the means ± SEM of percent changes over controls after normalization to the Renilla luciferase activity from three independent experiments in triplicate. Values with different letters vary statistically from each other (P<0.05) as determined by a one-way ANOVA followed by a Student-Newman-Keul’s test.

Qingli Shi, et al. J Neurochem. ;118(3):440-448.

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