QUICK SEARCH:   [advanced] Author: Keyword(s):
Home     Help     Feedback     Subscriptions     Archive     Search     Table of Contents

Help viewing high resolution images Return to article

Fig. 6. Hypoxia induces a persistent increase in RBM3 and CIRP protein levels via de novo mRNA synthesis. (A) HeLa cells were cultured in parallel in normoxia or 1% oxygen as indicated. Whole-cell lysates were isolated and subjected to western blot analysis for RBM3 and CIRP. To control sample loading and transfer, the blots were stripped and reprobed for ß-actin. (B) HeLa cells were cultured in parallel in normoxia or 1% oxygen exposed to actinomycin-D for 24 hours. RNA was isolated and subjected to real-time RT-PCR analysis. RBM3 and CIRP mRNA levels were normalized to ß₂-microglobulin (B2M) mRNA levels and are expressed as fold change in the different experiment samples compared with the corresponding normoxic samples (mean±s.e.m., n=3). (C) HeLa cells were cultured in parallel in normoxia or 1% oxygen as indicated for 24 hours. Nuclei were isolated, and in vitro transcription was allowed to resume for 40 minutes. RNA was isolated before and after in vitro transcription and subjected to real-time RT-PCR analysis. The extent of RBM3 and CIRP mRNA transcription was determined by subtracting the amount of RBM3 and CIRP mRNA standardized to B2M prior to transcription from the amounts post transcription (mean±s.e.m., n=3). Results are given as a ratio of copies of target gene (RBM3 or CIRP) to copies of the reference gene (B2M).

Return to article