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Figure 3 | Journal of Neuroinflammation

Figure 3

From: Hypothermia enhances induction of protective protein metallothionein under ischemia

Figure 3

Effects of metallothionein (MT) knock-down or overexpression on the cell death after ischemic insult. (A) We measured the lactate dehydrogenase (LDH) levels in culture media harvested from cells treated with siRNAs (10 nM) targeting MT-1 or -2 after 4 hours of oxygen glucose deprivation (OGD) and 24 hours of reperfusion at 33°C. *P <0.05 compared to 4 hours of OGD and 24 hours of reperfusion without siRNA treatment at 37°C; #P <0.05 compared to 4 hours of OGD and 24 hours of reperfusion without siRNA treatment at 33°C. Suppression of MT gene expression by siRNA was confirmed by measuring the level mRNA using RT-PCR. GAPDH was used as an internal standard. (B) We measured LDH levels in culture media harvested from the cells treated with MT overexpression vectors (2, 10, and 20 μg) after 4 hours of OGD and 24 hours of reperfusion at 37°C. Values represent the mean ± S.E.M. of three different experiments. *P <0.05 compared to 4 hours of OGD and 24 hours of reperfusion without MT overexpression vector at 37°C. Overexpression of the MT gene by the vector-containing human MT-1 clone was confirmed by detecting the levels of mRNA (RT-PCR) and protein (western blot) in different conditions: Without MT-1 vector and transfection reagent (CON), transfection reagent only (NC), MT-1 vector in transfection reagent (hMT). GAPDH was used as an internal standard.

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