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. 2012 Jun 28;287(34):29003–29020. doi: 10.1074/jbc.M112.373365

FIGURE 7.

FIGURE 7.

Effect of chronic hypoxia on Dicer mRNA and protein. A, relative RNA polymerase II (Pol II) occupancy at the Dicer proximal promoter in hypoxic HUVEC. Data represent mean ± S.E. (error bars) (n = 3). * denotes p < 0.05 compared with 0 h. B, Dicer pre-mRNA measurements in hypoxic HUVEC. Data represent mean ± S.E. (error bars) (n = 4). C, Dicer mRNA half-life measurements in control versus hypoxic HUVEC. Data represent mean ± S.E. (error bars) (n = 3). D, Dicer mRNA levels in nuclear (Nuc) versus cytoplasmic (Cyto) fractions of control versus 24-h hypoxic HUVEC. Data represent mean ± S.E. (error bars) (n = 3). * and † denote statistical significance (p < 0.05) compared with hypoxia-nuclear and control-cytoplasmic, respectively. Relative rate of global protein synthesis (E) and relative Dicer protein synthesis (F) in normoxic versus 24-h hypoxic HUVEC measured using [35S]methionine incorporation. A representative gel is shown for F. Data represent mean ± S.E. (error bars) (n = 3). * denotes p < 0.05 compared with normoxia. G, Dicer protein half-life measurements in control versus hypoxic HUVEC. Data represent mean ± S.E. (error bars) (n = 3). A representative gel from each condition is shown. H, representative immunoblots of control versus 24-h hypoxic HUVEC treated with MG-132 (left panel). Quantification of Dicer immunoblots is shown in the right panel. Data represent mean ± S.E. (error bars) (n = 3). * and † denote statistical significance (p < 0.05) compared with control-DMSO and hypoxia-DMSO, respectively. IP, immunoprecipitation.