Fig. 3. Plasminogen treatment of radiation wounds results in the decreased expression of genes involved in inflammation.

a–d, f Expression of selected inflammatory genes (as indicated) in radiation wounds before treatment (day 0) (n = 3) and at different time points of PBS (black) (n = 3) or plasminogen (red) (n = 3) treatment as measured with the Mouse Wound Healing RT² Profiler PCR array. The mRNA levels for all samples are presented as fold regulations relative to the mRNA levels in the control healthy, nonirradiated skin. (e) Quantification of 8-oxo-2-deoxyguanidine in healthy non-radiated skin (n = 3), radiated skin before treatment (day 0) (n = 3), and in PBS- (n ≥ 3) and plasminogen-treated (n ≥ 3) radiation wounds at days 20 and 30. g The KEGG enrichment scatter plot of inflammatory genes/pathways that were differentially expressed in plasminogen-treated wounds at day 20 when compared with PBS-treated wounds, as identified using mRNA sequencing. Downregulated pathways are shown in green, and the pathway that includes both upregulated and downregulated genes is shown in orange. h Expression of genes in the TLR pathway that were significantly changed at day 20 of plasminogen treatment when compared with PBS treatment, as measured using mRNA sequencing. i, j Quantification of protein levels for IL1-β and Spp1 (respectively) in healthy non-radiated skin, irradiated skin before treatment (day 0), and in PBS- and plasminogen-treated radiation wounds, based on immunostaining of paraffin sections (n ≥ 3). k Expression of genes in the TNF pathway that were significantly changed at day 20 of plasminogen treatment when compared with PBS treatment, as measured using mRNA sequencing. l Quantification of TNF-α protein levels as described in panel i (n ≥ 3). m Expression of genes in the arachidonic acid pathways that were significantly changed at day 20 of plasminogen treatment when compared with PBS treatment, as measured using mRNA sequencing. *P < 0.05; **P < 0.01; ***P < 0.005; ****P < 0.001. Rich factor is the ratio of the number of target genes divided by the number of all the genes in each pathway.