Journal: Oxidative Medicine and Cellular Longevity
Article Title: Hypoxia Downregulates MAPK/ERK but Not STAT3 Signaling in ROS-Dependent and HIF-1-Independent Manners in Mouse Embryonic Stem Cells
doi: 10.1155/2017/4386947
Figure Lengend Snippet: Relative expressions of DUSP1 (a), DUSP5 (b), and DUSP6 (c) in hypoxia in wild-type and HIF-1 α −/− ES cells. Statistical significance was determined by T test ( ∗ P < 0.05). Effect of siRNA-mediated silencing on DUSP1 (d), DUSP5 (e), and DUSP6 (f) expression in wild-type ES cells determined by qRT-PCR. Data are presented as mean + SEM from at least three independent experiments. Statistical significance was determined by one-way analysis of variance ANOVA and post hoc Bonferroni's multiple comparison test ( ∗ P < 0.05). Effect of DUSP1, 5, and 6 siRNAs on ERK phosphorylation and DUSP6 levels in wild-type ES cells (g). The total level of β -actin was used as a loading control. Densitometry analysis of western blots expressed as fold change in DUSP6 protein expression (h) or ERK phosphorylation (i). Data are presented as mean + SEM from at least three independent experiments. Statistical significance was determined by one-way analysis of variance ANOVA and post hoc Bonferroni's multiple comparison test ( ∗ P < 0.05).
Article Snippet: Cells were transfected by commercially available siRNA against DUSP1 (sc-35938), DUSP5 (sc-60555), DUSP6 (sc-39001) transcripts (each consisting of a pool of 3 target-specific 19-25 nt siRNAs designed to knock down gene expression), or related nonsilencing control (all Santa Cruz Biotechnology, USA) using Lipofectamine RNAiMAX Reagents (Thermo Fisher Scientific Inc., USA) according to the manufacturer's instructions.
Techniques: Expressing, Quantitative RT-PCR, Western Blot
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