text: Inhibition of transcription factors led to reduced IL-1RA levels
  Pharmacological inhibitors were used to demonstrate that the transcription factor sites within the IL-1RA promoter are relevant for the induction of IL-1RA by GP. As expected, an inhibition of NFkappaB via CAPE [35] and CyA, NFAT via CyA and NFIL-6 via CHX [36] could not be overruled by GP, leading to a down-regulated binding activity of the transcription factors to the IL-1RA sites, and a reduction of IL-1RA mRNA and IL-1RA protein levels (Fig. 7). The extent of reduction in NFATP2/3 binding is exemplarily shown in an autoradiogram (Fig. 7A, left side) and graphically summarized (n = 4, Fig. 7A, right side). The results for the NFkappaB and NFIL-6 binding sites were similar (data not shown). Because the highest IL-1RA amount was found at 24 h, mRNA was examined following 1 h inhibition and 18 h of GP. A representative gel demonstrating a decrease in IL-1RA mRNA after inhibition with CAPE, CyA and CHX is displayed in Fig. 7B (n = 4). In addition, corresponding IL-1RA protein levels after 24 h of GP are shown in Fig. 7C (n = 3). Altogether, NFkappaB inhibition by CAPE was more pronounced at the binding activity and the transcription, whereas CyA and CHX mainly led to a decreased IL-1RA release. 
  