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Figure 4 | Journal of Translational Medicine

Figure 4

From: Transcription and translation of human F11R gene are required for an initial step of atherogenesis induced by inflammatory cytokines

Figure 4

Upregulation of F11R mRNA expression by TNFα and INFγ in endothelial cells: inhibition by the NF-kB blocker and JAK protein kinase inhibitor. Panels (a) and (b). Confluent monolayers of HUVEC were pretreated (or Untreated) for 1 hr at 37°C with the NF-kB inhibitor, parthenolide (50 μM, final concentration), added to growth supplement-free media. The proinflammatory cytokines, TNFα (100 u/mL) or IFNγ (200 u/ml), were added to the media, and the cells were incubated at 37°C for an additional 24 hrs (see TNFα & Parthenolide in Figure 4a, and IFNγ & Parthenolide in Figure 4b). The response of cells exposed only to TNFα alone (100 u/ml) is shown in the histogram displayed in Figure 4a, and the response of cells exposed only to IFNγ alone is shown in Figure 4b. The F11R mRNA levels were measured by Real-time PCR performed in triplicate for each condition. Values are the mean ± SEM. * P < 0.05 level of significance observed between ECs exposed to TNFα or IFNγ alone vs ECs not exposed to TNFα/INFγ or ECs previously treated with parthenolide followed by their exposure to cytokines. Figure 4c demonstrates the upregulation of F11R mRNA in endothelial cells by IFNγ and its inhibition by the JAK protein kinase inhibitor, AG-490. Confluent monolayers of HUVEC were either Untreated or treated with the JAK protein kinase inhibitor AG-490 (50 μM) alone (AG 490) added to growth supplement-free media and incubated for 1 hr at 37°C. The response of cells that were exposed to the cytokine IFNγ alone is depicted in the histogram IFNγ. The response of cells that were pretreated with AG 490 for 1 hr followed by their exposure to IFNγ (200 u/mL) for an additional 24 hrs is depicted in histogram labeled IFNγ & AG-490. The F11R mRNA levels were measured by Real-time PCR performed in three separate experiments, in triplicate, for each condition. Values are the mean ± SEM. * P < 0.05 significance differences in F11R mRNA in ECs exposed to IFNγ alone vs untreated ECs or ECs treated with AG-490 alone or ECs previously treated with AG-490 followed by their exposure to IFNγ

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