Temporal and spatial expression profiles of sample genes from the three inflammation-dependent clusters. Temporal and spatial profiles of the (A) early inflammatory, (B) late inflammatory and (C) inflammation-maintained clusters. Line graphs display absolute temporal expression levels (y-axis) at each time point (x-axis) for both PU.1 null (pink) and wild-type (blue) wounds. y-axis expression levels vary according to individual gene expression levels. In situ hybridization studies of (A) 12 h, (B) 24 h or (C) 3 h frozen sections illustrate the contrasting expression patterns of each of these classes of genes in wild-type (WT) versus PU.1 null wounds. (Aa,d,g,j,m,p,s) In the early inflammatory cluster, expression in wild-type wounds peaks at 12 h but is absent or significantly reduced in PU.1 null wounds. (Ab,c) In situ studies show L-plastin to be expressed by activated leukocytes in the wild-type only (arrow). (Ae,f) Faint expression of C3 is seen in both genotypes (see arrows). (Ah,i) Onzin expression appears to be in the same cells within the connective tissue in both genotypes. (Ak,l) Both keratinocytes and leukocytes (arrows) express MRP14 in the wild-type but only keratinocyte expression (arrow) is seen in the PU.1 null wound. (An,o) Osteopontin displays a possible 'fibrosis' gene spatial profile with expression in deep dermal cell layers (arrow) in the wild-type only. (Aq,r) CCr1 is expressed only in the wild-type wound, in cells whose clustered location suggests they are one of the leukocyte lineages.(At,u) Expression of CXC10 is broad and throughout the wound connective tissue of wild-type wounds (arrow) suggesting that expressing cells are wound fibroblasts. (B) In the late inflammatory cluster, expression in wild-type wounds appears to peak beyond 12 h in wild-type wounds and is absent or reduced in PU.1 null wounds. (Bb,c) Expression of Cathepsin S is seen in activated leukocytes in the wild-type only (arrow). (Bd) Repetin is expressed by both genotypes but to a lower level in the PU.1 null. (Be,f) Repetin is only upregulated by keratinocytes but is not restricted only to wild-type wounds (arrows). (Bh,i) Expression of the potential fibrosis gene Angiotensin II Receptor 1 is seen in deep dermal cell layers of wild-type and, to a significantly reduced level, PU.1 null wounds (arrows). (C) In the inflammation maintained cluster, the expression profiles suggest that while these genes may be initially expressed in PU.1 null wounds, persistent expression requires the presence of an inflammatory response as in the wild-type wound situation. (Cb,c) Expression of Mcpt5 is seen at both wound sites (arrows) in scattered cells throughout the wound connective tissue. (Ce,f) CCL2 appears to be expressed by host wound cells at both wound sites (see arrows). (Ch,i) CCL7 is expressed in an almost identical temporal and spatial profile to CCL2. Scale bars = 400 μm (Aa-o, Ba-f, C) and 250 μm (Ap-u and Bh,i).
Cooper et al. Genome Biology 2004 6:R5 doi:10.1186/gb-2004-6-1-r5