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Fig. 8. Induction of RBM3 and CIRP in response to hypoxia does not require mitochondria. mtDNA was depleted from HeLa cells with EB in two different concentrations for 6 days as indicated. Subsequently cells were cultured in parallel in normoxia or 1% oxygen as indicated for 24 hours. Parental (WT) and rho0 143B cells, which are devoid of mtDNA, were used as control. Total DNA and RNA was isolated. The upper diagram shows the relative percentage (amount relative to the untreated control cell lines) of mtDNA, assessed by quantitative real-time PCR (see Materials and Methods). RNA was subjected to real-time RT-PCR analysis; RBM3, CIRP and VEGF RNA levels were quantitated, normalized to B2M RNA levels and are expressed as fold change in the different experiment samples compared with the corresponding normoxic samples (mean±s.e.m., n=3).
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