Inhibition of N-Myc down regulated gene 1 in in vitro cultured human glioblastoma cells

Figure 3 Inhibition of N-Myc down-regulated gene 1 protein and mRNA expression in U373 human glioblastoma cell line in vitro via short dsRNA oligonucleotides- and iodoacetate-mediated interference into human tumor cellular glycolysis process. Upper panel: Western blotting analysis result diagram related to the specific inhibition of hypoxia induced expression in human brain tumor cells (U373 Glioblastoma cell line as an example) on protein level via Western blotting analysis. Clear complete inhibition of N-Myc down-regulated gene 1 (NDRG1) induced by extreme hypoxic conditions in the tumor microenviroment (0.1% O₂/for 24 h) was present upon transfection with the one of the two variants of NDRG1 short dsRNA oligonucleotides (siRNA) construct applied in these experiments when compared to the nearly complete inhibition via inhibitive interaction with the tumor cell glycolysis pathway. β-actin served as a loading control. Figure shows one representative experiment out of three experiments; Lower pannel: Northern blotting analysis displaying the specific inhibition of hypoxia induced NDRG1 mRNA expression in human brain tumor cells (U373 Glioblastoma cell line as an example) on mRNA level. Complete inhibition of NDRG1 induced by extreme hypoxic conditions in the tumor microenviroment (0.1% O₂/for 24 h) was could be achieved upon transfection with the one of the two variants of NDRG1 siRNA construct applied in these experiments. Inhibitive interaction into the glycolysis pathway due to the parallel treatment with 50 μmol/L with iodoacetate for 24 h showed a similar functional effect. The 18s RNA fragment with a molecular weight of 1.9 kb served as a loading control. This is one representative experiment out of three experiments.