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Fig. 3. Regulation of HERC5 mRNA expression. (A) HERC5 gene expression in HM2 cells in response to different stimuli after 8 hours of treatment. 10 µg of total RNA were loaded per lane for analysis. Northern blots were probed for HERC5 expression and reprobed for GAPDH to confirm equal loading of RNA. CHX, cycloheximide; EGF, epidermal growth factor; IL, interleukin; LPS, lipopolysaccharide; TGFß, transforming growth factor ß; TNF{alpha}, tumor necrosis factor {alpha}, VEGF, vascular endothelial growth factor. (B) Northern blot analysis of rAd.I{kappa}B{alpha}-infected cells. HSMEC were infected with either the rAd. I{kappa}B{alpha} or a control adenovirus as indicated above each lane and 48 hours post infection cells were left unstimulated or stimulated with LPS for 6 hours. 10 µg of total RNA were analyzed for mRNA of HERC5, I{kappa}B{alpha} and GAPDH by subsequent hybridizations. (C) HUMECs were treated as described in B but stimulated with LPS for 8 hours; isolated RNA was analyzed by quantitative real-time RT-PCR. *P<0.05.





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