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. 2004 Oct 4;101(41):14871–14876. doi: 10.1073/pnas.0406073101

Fig. 4.

Fig. 4.

Athero-prone waveform stimulation results in enhanced IL-8 production and NF-κB nuclear translocation. (a) IL-8 mRNA expression was measured by TaqMan RT-PCR (n = 3) at 24 h. All data are normalized to the static (no flow) condition. (b) Endothelial IL-8 secretion was measured by ELISA (n = 3). EC were cultured under static (no flow) or athero-prone and athero-protective waveform for 24 h. The supernant was collected from the outflow port of the dynamic flow device every 2 h, and IL-8 protein was measured by ELISA (see Materials and Methods). (c) Immunofluorescent staining of NF-κB (p65) (×40). EC were stained with antibodies to p65 after 24 h exposure to static (no flow) and athero-prone and athero-protective waveform. (d) Western blot analysis of NF-κB (p65) protein levels. HUVEC were cultured under static, athero-prone, or athero-protective flow conditions for 24 h, then treated with IL-1β (1 units/ml) for 1 h under static condition. Cytoplasmic and nuclear protein fractions were isolated and p65 protein was analyzed by Western blotting. The blot was then stripped and reprobed for α-tubulin and histone H3. Lanes 1, static (no flow); lanes 2, athero-prone; lanes 3, athero-protective condition.