Transient transfection analysis of an SGP promoter (NC) harboring a Gfi-1 and a C/EBP site. (A) DNA sequence of the promoter of an SGP gene, neutrophil collagenase (NC). The first 193 bp of the human (HNC193) neutrophil collagenase promoter sequences representing the minimal promoter is illustrated. The 3 C/EBP sites (P, I, and D) are underlined in black and the conserved Gfi-1 sequence, in gray. The transcription start site is marked by a +1 sign and the translation start site ATG is indicated. (B) 32Dwt18 cells were transiently cotransfected with wild-type HNC193 (wt) or Gfi-1 mutant HNC193 (“Patients, materials, and methods”) and 10 μg expression plasmids for Gfi-1 and wt C/EBPϵ (e) separately or together. HNC193 (wt) was also cotransfected with the Gfi-1 expression plasmid and wt or V218A C/EBPϵ plasmids. Normalized luciferase values have been represented as a ratio of enzyme activity of HNC193 promoter plasmid plus C/EBP and/or Gfi-1 expression plasmids to that of HNC193 promoter plasmid without expression plasmids. The figure represents normalized mean ± SE obtained from 3 independent experiments, each performed in duplicate. Statistical significance was determined using Student t test, and the P values have been indicated.