Fig. 1

Map of SCD1 promoter in cattle and determination of SCD1 expression and chromatin compaction. a The tsp as identified in 5′-RACE experiments is indicated (+1; hatched box, exon 1) and positions of selected putative transcription factor bindings sites (in silico analysis; symbols explained in legend below) are mapped relative to the tss. Encircled is a doubled site of overlapping C/EBP and NF-κB binding sites, at around position −1020. Short kinked arrows indicate the 5′-ends of the long and short promoter segments (PL, PS, respectively) used in reporter gene assays. Nucleotide sequences of two C/EBP binding sites are given below the map (core nucleotides in bold face letters). The mutated variants hereof are shown below (sequence alterations underlined). Gray bars label the three areas (A, B, C) having been analyzed in CHART-PCR exploiting the restriction enzyme cutting sites as indicated by the vertical arrows. b The degree of chromatin compaction in three different promoter areas (areas A, B, C), as determined in CHART-PCR (abscissa, expressed as percent of undigested control) is plotted against the levels of SCD1 mRNA concentration (ordinate) from healthy control cows (grey squares) or cows having experimentally been infected for 24 h with E. coli (diamonds). A Distal area A, analyzed with primers pfA/prA and ScaI digestion; B Area around the SREBP1 binding site, analyzed with primers pfB/prB and MspI digestion. C Proximal area C, analyzed with primers pfC/prC and MspI digestion. The degree of chromatin compaction did not significantly differ between livers from both groups of cows and there was no correlation between chromatin compaction and the level of gene expression