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Figure 2


Fig. 2. Regulation of HBEGF accumulation by O2. (A) Western blot of HBEGF. Recombinant human HBEGF (20 ng in lane 1) contains the same sequence as sHBEGF. HBEGF was undetectable in conditioned medium (lane 2) or in cytotrophoblast cell lysate (lane 3) during culture at ambient O2 levels. After 24 hours at 2% O2, a 9.7 kDa sHBEGF band was detected in the medium (lane 4), and an 18.7 kDa pro-HBEGF band appeared in cell lysates in addition to the sHBEGF band (lane 5). (B) Cellular and secreted pools of HBEGF were quantified by ELISA in cell lysates and conditioned medium after shifting cells to 2% O2. *P<0.05, compared with control (0 hours). (C) Northern blot of the 2.5 kb HBEGF transcript in cytotrophoblast RNA 0-24 hours after shifting cells to 2% O2. (D) Real time RT-PCR was used to quantify HBEGF mRNA 0-24 hours after shifting cells to 2% O2. ANOVA indicated no significant differences.





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