(A) The enhancer DNA corresponding to eRNA 134 was subcloned downstream of luciferase under the control of the minimal thymidine kinase promoter (TK). The firefly luciferase vectors were co-transfected with a constitutive Renilla control, pRL-TK, into HEK293 cells. PGL3-TK-A7 is a positive control from near the UBE2VI gene. Construct 172 is a negative control, which encodes a non-coding RNA but is not an eRNA and is not located in the ADAM locus. Inducible firefly luciferase was normalized to the Renilla signal. DNA from Enhancer 1 was able to transactivate TK expression in this system. (B) The Enhancer 1 and 2 eRNAs were coordinately regulated with ADAMDEC1 in primary monocytes. The cells were pretreated by SB203580, JSH-23, SP60025 or U0126 for 30 minutes and then stimulated with LPS for 90 minutes (n=3, error bars in panels denote SD). Symbols indicate statistical significance: * indicates p<0.05, ** p<0.01, and *** p<0.001.